Novel compound having blt inhibitory activity and composition, for preventing or treating inflammatory diseases, comprising same as active ingredient

ABSTRACT

The present invention relates to a novel compound showing leukotriene B4 receptor 2 (BLT2) inhibitory activity and a pharmaceutical composition, for preventing or treating inflammatory diseases, having same as an active ingredient. The inventors identified a novel compound containing BTL2 inhibitory activity, and experimentally confirmed that the present novel compound had an excellent effect on the enhancement of the cancer cell death, on the inhibition of the metastasis and chemotactic mobility, and on the anti-asthma activity. Therefore, the present novel compound can be used as a very effective pharmaceutical component for treating the inflammatory-related diseases.

CROSS-REFERENCE TO RELATED APPLICATIONS

This Application is a Divisional of U.S. application Ser. No.15/745,337, filed Mar. 30, 2018, Which in turn is 371 ofPCT/KR2016/008069, filed Jul. 23, 2016, which claims the benefit ofpriority from Korean Patent Application No. 10-2015-0105097, filed Jul.24, 2015 and Korean Patent Application No. 10-2016-0093762, filed Jul.22, 2016, the contents of each of which are incorporated herein byreference.

TECHNICAL FIELD

The present invention relates to a novel compound and a use thereof, andmore particularly, to a novel compound exhibiting a leukotriene B4receptor 2 (BLT2) inhibitory activity and a pharmaceutical compositionfor preventing or treating an inflammatory disease, which includes thenovel compound as an active ingredient.

BACKGROUND ART

An inflammatory response is one of the human immune systems activated byvarious action mechanisms to defend against physical actions, chemicals,bacterial infections, or immunological stimuli, which are applied toliving organisms or tissue. However, when such inflammatory responsepersists, rather, damage to a mucous membrane is promoted, and thereforeit has been noted that inflammatory diseases including rheumatoidarthritis, atherosclerosis, gastritis, asthma, etc. are caused byerythema, a fever, swelling, pain, or dysfunction. Such an inflammatoryresponse is classified into acute inflammation and chronic inflammationas time passes. The acute inflammation is an inflammatory responselasting several days to several weeks, and causes a symptom such aserythema, a fever, pain, or swelling, whereas the chronic inflammationis a long-term inflammatory state for several years to decades, andinvolves a histological change such as fibrosis or the destruction oftissue caused by the infiltration of monocytes, proliferation offibroblasts or capillaries, or an increase in connective tissue.

Specifically, when inflammatory stimuli are applied to the livingorganism, locally, histamine, bradykinin, prostaglandins, nitric oxide(NO), all types of pro-inflammatory cytokines, etc. are synthesized andsecreted, and cause erythema, a fever, pain, or swelling as well asvasodilation. Particularly, in inflammation in the body, in addition tocommon immune factors, for example, cytokines such as interferon-γ(INF-γ), tumor necrosis factor-α (TNF-α), interleukin-1 (IL-1), andinterleukin-6 (IL-6), nitric oxide (NO) and prostaglandin E2 (PGE2) arewell known as major proinflammatory materials.

Conventionally, the termination of an inflammation response is known asa phenomenon naturally and passively occurring due to a decrease inlevels of materials initiating inflammation, but it was found that thetermination of inflammation is actively promoted by lipoxins, resolvins,or protectins, which were discovered by Serhan et al., likeprostaglandins, which are involved in the initiation of inflammation.For example, it has been reported that Resolvin E1 is effective forpain, and RvE1 induces the termination of inflammation and is effectivein treating an allergic inflammatory disease. In addition, it has beenreported that low levels of factors actively promoting the terminationof such inflammation in a chronic inflammatory disease, that is, lipoxinA4 and lipoxin induced by aspirin are shown in asthmatic patients andatherosclerotic patients.

Accordingly, while various attempts to find novel materials for inducingthe termination of inflammation and thus to treat diseases associatedwith abnormal inflammation termination have been made (Korean UnexaminedPatent Application No. 10-2015-0011875), a compound known to be includedin lipoxins, resolvins, etc. is metabolically unstable and thus rapidlydegraded in the body due to several double bonds in its structure, andis somewhat difficult to be developed as a drug by mass production of amaterial, thereby having a great problem in drugability.

Meanwhile, leukotriene B4 (LTB₄) is a group of inflammatory lipidmediators synthesized from arachidonic acid (AA) via a 5-lipoxygenasepathway mediating acute and chronic inflammation. LTB₄ is known to givea biological effect by binding to two types of receptors such as BLT1and BLT2. Leukotriene B4 receptor 2 (BLT2), as one among the Gprotein-coupled receptor (GPCR) family, is a receptor having lowaffinity to LTB₄, and a lipid mediator of arachidonic acid (AA) inducedvia a 5-lipoxygenase-dependent pathway.

Accordingly, to solve the above-mentioned conventional problems, theinventors prepared a novel compound exhibiting a BLT2 inhibitoryactivity while conducting research to develop a material for inducingthe effective termination of inflammation, and first invented atherapeutic agent for an inflammatory disease, which includes theabove-mentioned compound.

DISCLOSURE Technical Problem

The present invention is provided to solve the above-mentioned problems,and the inventors confirmed a therapeutic effect of a novel compoundexhibiting a BLT2 inhibitory activity on an inflammatory disease, andbased on this, the present invention was completed.

Therefore, an object of the present invention is to provide a novelcompound exhibiting a BLT2 inhibitory activity or a pharmaceuticallyacceptable salt thereof.

Another object of the present invention is to provide a pharmaceuticalcomposition for preventing or treating an inflammatory disease, whichincludes the novel compound or pharmaceutically acceptable salt thereofas an active ingredient.

However, technical problems to be solved in the present invention arenot limited to the above-described problems, and other problems whichare not described herein will be fully understood by those of ordinaryskill in the art from the following description.

Technical Solution

To achieve these objects of the present invention, the present inventionprovides a novel compound exhibiting a BLT2 inhibitory activity or apharmaceutically acceptable salt thereof.

According to an exemplary embodiment of the present invention, thecompound may be selected from the group consisting of tert-butyl4-(4-(3-(N-phenylpentaneamido)prop-1-ynyl)benzoyl)piperazine-1-carboxylate;N-phenyl-N-(3-(4-(piperazine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide;N-(3-(4-(4-methylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentaneamide;N-(3-(4-(4-ethylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentaneamide;N-(3-(4-(4-isopropylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentaneamide;N-(3-(4-(4-(2-hydroxyethyl)piperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentaneamide;N-(3-(4-(4-(cyclopropylmethyl)piperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentaneamide;N-(3-(4-(4-cyclohexylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentaneamide;N-(3-(4-(4-(cyclohexylmethyl)piperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentaneamide;N-(3-(4-(4-isobutylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentaneamide;N-phenyl-N-(3-(4-(4-(prop-2-ynyl)piperazine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide;N-(3-(4-(4-cyanopiperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentaneamide;tert-butyl4-(4-(3-(N-(3-fluorophenyl)pentaneamido)prop-1-ynyl)benzoyl)piperazine-1-carboxylate;N-(3-fluorophenyl)-N-(3-(4-(piperazine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide;N-(3-fluorophenyl)-N-(3-(4-(4-isopropylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide;tert-butyl4-(4-(3-(N-(4-fluorophenyl)pentaneamido)prop-1-ynyl)benzoyl)piperazine-1-carboxylate;N-(4-fluorophenyl)-N-(3-(4-(piperazine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide;N-(4-fluorophenyl)-N-(3-(4-(4-isopropylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide;N-(3-(4-(morpholine-4-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentaneamide;N-phenyl-N-(3-(4-(piperidine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide;N,N-diethyl-4-(3-(N-phenylpentaneamido)prop-1-ynyl)benzamide;N-phenyl-N-(3-(3-(piperazine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide;N-(3-(3-(4-methylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentaneamide;N-(3-(3-(4-isopropylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentaneamide;tert-butyl-4-(3-(3-(N-(4-fluorophenyl)pentaneamido)prop-1-ynyl)benzoyl)piperazine-1-carboxylate;N-(4-fluorophenyl)-N-(3-(3-(piperazine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide;N-(4-fluorophenyl)-N-(3-(3-(4-isopropylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide;N-(3-(4-hydroxyphenyl)prop-2-ynyl)-N-phenylpentaneamide;2-(4-(3-(N-phenylpentaneamido)prop-1-ynyl)phenoxy)acetic acid;tert-butyl4-(5-(3-((N-phenylpentaneamido)prop-1-yn-1-yl)picolinoyl)piperazine-1-carboxylate;N-phenyl-N-(3-(6-(piperazine-1-carbonyl)pyridine-3-yl)prop-2-yn-1-yl)pentaneamide;N-(3-(6-isopropylpiperazine-1-carbonyl)pyridine-3-yl)prop-2-yn-1-yl)pentaneamide;N,N-diethyl-4-(3-(N-(3-fluorophenyl)pentamido)prop-1-yn-1-yl)benzamide;N,N-diethyl-4-(3-(N-(4-fluorophenyl)pentamido)prop-1-yn-1-yl)benzamide;N-(3-(4-(N,N-diethylsulfamoyl)phenyl)prop-2-ynyl)-N-phenylpentamide;N-(3-(4-(N-isopropylsulfamoyl)phenyl)prop-2-ynyl)-N-phenylpentamide;tert-butyl 4-(3-(N-phenylpentaneamido)prop-1-yn-1-yl)benzoate;4-(3-(N-phenylpentaneamido)pro-1-yn-1-yl)benzoic acid;N-ethyl-4-(3-(N-phenylpentaneamido)prop-1-yn-1-yl)benzamide;N-(2-(diethylamino)ethyl)-4-(3-(N-phenylpentaneamido)prop-1-yn-1-yl)benzamide;ethyl 2-(4-(3-(N-phenylpentaneamido)prop-1-yn-1-yl)benzamido)acetate;2-(4-(3-(N-phenylpentaneamido)prop-1-yn-1-yl)benzamido)acetic acid;methyl2-(4-(3-(N-phenylpentaneamido)prop-1-yn-1-yl)benzamido)propanoate;2-(4-(3-(N-phenylpentaneamido)prop-1-yn-1-yl)benzamido)propionic acid;2-(4-(3-(N-(3-fluorophenyl)pentaneamido)prop-1-ynyl)phenoxy)acetic acid;and 2-(4-(3-(N-(4-fluorophenyl)pentaneamido)prop-1-ynyl)phenoxy)aceticacid.

The present invention provides a pharmaceutical composition forpreventing or treating an inflammatory disease, which includes the novelcompound or a pharmaceutically acceptable salt thereof as an activeingredient.

According to an exemplary embodiment of the present invention, theinflammatory disease may be selected from the group consisting ofasthma, atherosclerosis, cancer, pruritus, rheumatoid arthritis andinflammatory enteropathy.

According to another exemplary embodiment of the present invention, thecomposition may inhibit BLT2 activity.

The present invention provides a method for treating an inflammatorydisease, which includes administering the pharmaceutical composition toa subject.

The present invention provides a use of the composition including thenovel compound or a pharmaceutically acceptable salt thereof to treat aninflammatory disease.

Advantageous Effects

The present invention relates to a new compound exhibiting BLT2inhibitory activity and a pharmaceutical composition for preventing ortreating an inflammatory disease, which includes the compound. Theinventors identified a novel compound exhibiting BTL2 inhibitoryactivity to solve problems of a conventional material for treating aninflammatory disease, for example, instability in the living organismand difficulty in mass production, and experimentally confirmed that thecompound has excellent effects of improving the death of cancer cellsand inhibiting cancer cell metastasis, a chemotactic motility inhibitoryeffect, and an antiasthma effect, and therefore the compound is expectedto be effectively used as a pharmaceutical composition for treating aninflammatory disease.

DESCRIPTION OF DRAWINGS

FIGS. 1A to 1E show the results of confirming a growth inhibitory effectin BLT2-expressing cells (CHO-BLT2 cells) caused by treatment of acompound of the present invention.

FIGS. 2A and 2B show the results of confirming an effect of inhibitingcell chemotactic motility and the 50% inhibition concentration (IC₅₀) inBLT2-expressing cells (CHO-BLT2 cells) by treatment of a compound of thepresent invention.

FIGS. 3A and 3B show the results of confirming an effect of inhibitingcell chemotactic motility in BLT2-expressing cells (CHO-BLT2 cells) orBLT1-expressing cells (CHO-BLT1 cells) by treatment of a compound of thepresent invention.

FIGS. 4A and 4B show the results of confirming LTB₄ and BLT2 bindinginhibitory effects in BLT2-expressing cells (CHO-BLT2 cells) bytreatment of a compound of the present invention.

FIGS. 5A and 5B show the results of confirming an effect of inhibitingthe generation of reactive oxygen species in MDA-MB-231 cells orMDA-MB-435 cells by treatment of a compound of the present invention.

FIGS. 6A and 6B show the results of confirming an effect of inhibitinginterleukin-8 (IL-8) expression levels in MDA-MB-231 cells or MDA-MB-435cells by treatment of a compound of the present invention.

FIGS. 7A and 7B show the results of confirming an effect of inhibitingcancer cell invasion in MDA-MB-231 cells or MDA-MB-435 cells bytreatment of a compound of the present invention.

FIGS. 8, 9A and 9B show the results of confirming an effect ofinhibiting cancer cell metastasis by treatment of a compound of thepresent invention.

FIG. 10 shows the results of confirming an effect of reducing airwayhyperresponsiveness (AHR) in severe asthma-induced mice by treatment ofa compound of the present invention.

FIG. 11 shows the results of confirming an effect of reducinginterleukin-4 (IL-4) generation in severe asthma-induced mice bytreatment of a compound of the present invention.

FIG. 12 shows the results of confirming an effect of reducing airwayhyperresponsiveness (AHR) in severe asthma-induced mice by treatment ofa compound of the present invention.

FIGS. 13A to 13C show the results of confirming that the influx of totalcells and neutrophils into a mouse abdominal cavity is reduced inasthma-induced mice by treatment of a compound of the present invention.

FIGS. 14A and 14B show the results of confirming that the influx oftotal cells and neutrophils into a mouse abdominal cavity inasthma-induced mice is reduced by treatment of a compound of the presentinvention.

MODES OF THE INVENTION

The inventors specifically identified effects of improving the death ofcancer cells, the inhibition of cancer cell metastasis and theinhibition of BLT2-dependent chemotactic motility, and an antiasthmaeffect based on the fact that the growth of BLT2-expressing cells can beconsiderably inhibited when a novel compound prepared in an example istreated, and therefore, the present invention was completed.

Hereinafter, the present invention will be described in detail.

The present invention provides a compound represented by Formula 1 belowor a pharmaceutically acceptable salt thereof.

In Formula 1,

R₁ may be a C₁ to C₁₀ alkyl,

R₂ may be

R_(a) may be

or hydroxy,

R_(b) may be

R_(c) may be

R_(d) may be hydrogen or

R_(e) may be

and

R₃ may be hydrogen or fluorine.

Exemplary examples of the compound represented by Formula 1 according tothe present invention are as follows: tert-butyl4-(4-(3-(N-phenylpentaneamido)prop-1-ynyl)benzoyl)piperazine-1-carboxylate;N-phenyl-N-(3-(4-(piperazine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide;N-(3-(4-(4-methylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentaneamide;N-(3-(4-(4-ethylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentaneamide;N-(3-(4-(4-isopropylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentaneamide;N-(3-(4-(4-(2-hydroxyethyl)piperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentaneamide;N-(3-(4-(4-(cyclopropylmethyl)piperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentaneamide;N-(3-(4-(4-cyclohexylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentaneamide;N-(3-(4-(4-(cyclohexylmethyl)piperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentaneamide;N-(3-(4-(4-isobutylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentaneamide;N-phenyl-N-(3-(4-(4-(prop-2-ynyl)piperazine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide;N-(3-(4-(4-cyanopiperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentaneamide;tert-butyl4-(4-(3-(N-(3-fluorophenyl)pentaneamido)prop-1-ynyl)benzoyl)piperazine-1-carboxylate;N-(3-fluorophenyl)-N-(3-(4-(piperazine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide;N-(3-fluorophenyl)-N-(3-(4-(4-isopropylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide;tert-butyl4-(4-(3-(N-(4-fluorophenyl)pentaneamido)prop-1-ynyl)benzoyl)piperazine-1-carboxylate;N-(4-fluorophenyl)-N-(3-(4-(piperazine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide;N-(4-fluorophenyl)-N-(3-(4-(4-isopropylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide;N-(3-(4-(morpholine-4-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentaneamide;N-phenyl-N-(3-(4-(piperidine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide;N,N-diethyl-4-(3-(N-phenylpentaneamido)prop-1-ynyl)benzamide;N-phenyl-N-(3-(3-(piperazine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide;N-(3-(3-(4-methylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentaneamide;N-(3-(3-(4-isopropylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentaneamide;tert-butyl-4-(3-(3-(N-(4-fluorophenyl)pentaneamido)prop-1-ynyl)benzoyl)piperazine-1-carboxylate;N-(4-fluorophenyl)-N-(3-(3-(piperazine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide;N-(4-fluorophenyl)-N-(3-(3-(4-isopropylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide;N-(3-(4-hydroxyphenyl)prop-2-ynyl)-N-phenylpentaneamide;2-(4-(3-(N-phenylpentaneamido)prop-1-ynyl)phenoxy)acetic acid;tert-butyl4-(5-(3-(N-phenylpentaneamido)prop-1-yn-1-yl)picolinoyl)piperazine-1-carboxylate;N-phenyl-N-(3-(6-(piperazine-1-carbonyl)pyridine-3-yl)prop-2-yn-1-yl)pentaneamide;N-(3-(6-isopropylpiperazine-1-carbonyl)pyridine-3-yl)prop-2-yn-1-yl)pentaneamide;N,N-diethyl-4-(3-(N-(3-fluorophenyl)pentamido)prop-1-yn-1-yl)benzamide;N,N-diethyl-4-(3-(N-(4-fluorophenyl)pentamido)prop-1-yn-1-yl)benzamide;N-(3-(4-(N,N-diethylsulfamoyl)phenyl)prop-2-ynyl)-N-phenylpentamide;N-(3-(4-(N-isopropylsulfamoyl)phenyl)prop-2-ynyl)-N-phenylpentamide;tert-butyl 4-(3-(N-phenylpentaneamido)prop-1-yn-1-yl)benzoate;4-(3-(N-phenylpentaneamido)pro-1-yn-1-yl)benzoic acid;N-ethyl-4-(3-(N-phenylpentaneamido)prop-1-yn-1-yl)benzamide;N-(2-(diethylamino)ethyl)-4-(3-(N-phenylpentaneamido)prop-1-yn-1-yl)benzamide;ethyl 2-(4-(3-(N-phenylpentaneamido)prop-1-yn-1-yl)benzamido)acetate;2-(4-(3-(N-phenylpentaneamido)prop-1-yn-1-yl)benzamido)acetic acid;methyl2-(4-(3-(N-phenylpentaneamido)prop-1-yn-1-yl)benzamido)propanoate;2-(4-(3-(N-phenylpentaneamido)prop-1-yn-1-yl)benzamido)propionic acid;2-(4-(3-(N-(3-fluorophenyl)pentaneamido)prop-1-ynyl)phenoxy)acetic acid;and 2-(4-(3-(N-(4-fluorophenyl)pentaneamido)prop-1-ynyl)phenoxy)aceticacid.

The term “pharmaceutically acceptable” used herein refers to a compoundor composition that is suitable to be used in contact with a subject's(e.g., a human) tissue due to a reasonable benefit/risk ratio withoutexcessive toxicity, irritation, allergic reactions, or other problems orcomplications, and included within the scope of sound medical judgment.

The term “salt” used herein is an acid addition salt formed by apharmaceutically acceptable free acid. The acid addition salt isobtained from inorganic acids such as hydrochloric acid, nitric acid,phosphoric acid, sulfuric acid, hydrogen bromide, hydrogen iodide,nitride and phosphorous acid, and non-toxic organic acids such asaliphatic mono and dicarboxylates, phenyl-substituted alkanoates,hydroxyl alkanoates and alkandioates, aromatic acids, aliphatic andaromatic sulfonic acids. Such pharmaceutically non-toxic salts includesulfates, pyrosulfates, bisulfates, sulfites, bisulfites, nitrates,phosphates, monohydrogen phosphates, dihydrogen phosphates,metaphosphates, pyrophosphate chlorides, bromides, iodides, fluorides,acetates, propionates, decanoates, caprylates, acrylates, formates,isobutyrates, caprates, heptanoates, propiolates, oxalates, malonates,succinates, suberates, sebacates, fumarates, maleates,butyne-1,4-dioates, hexane-1,6-dioates, benzoates, chlorobenzoates,methylbenzoates, dinitrobenzoates, hydroxylbenzoates, methoxybenzoates,phthalates, terephthalates, benzenesulfonates, toluenesulfonates,chlorobenzenesulfonates, xylenesulfonates, phenylacetates,phenylpropionates, phenylburyrates, citrates, lactates,β-hydroxylbutyrates, glycolates, malates, tartrates, methanesulfonates,propanesulfonates, naphthalene-1-sulfonates, naphthalene-2-sulfonates,and mandelates.

The acid addition salt according to the present invention may beprepared by a conventional method, for example, dissolving compoundsrepresented by Formulas 1 to 4 in an excessive acid aqueous solution,and precipitating the resulting salt using a water-miscible organicsolvent, for example, methanol, ethanol, acetone or acetonitrile. Inaddition, the acid addition salt according to the present invention maybe prepared by evaporating a solvent or an excessive acid from thismixture, and then dehydrating the resulting mixture orsuction-filtrating a precipitated salt.

In addition, the pharmaceutically acceptable metal salt may be preparedusing a base. An alkali metal or alkali earth metal salt may be obtainedby, for example, dissolving a compound in an excessive amount of analkali metal hydroxide or alkali earth metal hydroxide solution,filtering an insoluble compound salt, and dehydrating the remainingsolution through evaporation. Here, a sodium, potassium or calcium saltis pharmaceutically appropriate for the metal salt. Also, a silver saltcorresponding to the metal salt is obtained by a reaction between analkali metal or alkali earth metal salt and a suitable silver salt(e.g., silver nitrate).

In an exemplary embodiment of the present invention, novel compoundsexhibiting a BLT2 inhibitory activity were prepared (see Examples 1 to46), and it was confirmed that the growth of BLT2-expressing cells wereinhibited by the treatment of the novel compound (see ExperimentalExample 2). In addition, it was confirmed that chemotactic motility ofthe BLT2-expressing cells can be inhibited (see Experimental Example 3).In addition, an LTB₄ and BLT2 binding inhibitory effect was confirmedusing the compound (see Experimental Example 4), the inhibition ofreactive oxygen species in cells, the inhibition of IL-8 expression, theinhibition of cancer cell invasion, and the inhibition of cancer cellmetastasis were confirmed (see Experimental Example 5), and it was alsospecifically confirmed that the compounds have effects of reducingairway hyperresponsiveness (AHR), inhibiting IL-4 generation andinhibiting the influx of immune cells into a mouse abdominal cavity inasthma-induced mice (see Experimental Example 6), and therefore it wasconfirmed that the compounds can be very effectively used aspharmaceutical composition for an inflammatory disease.

Thus, the present invention provides a pharmaceutical composition forpreventing or treating an inflammatory disease, which includes thecompound or a pharmaceutically acceptable salt thereof.

Meanwhile, the term “prevention” used herein refers to all actions ofinhibiting an inflammatory disease or delaying the onset thereof byadministration of the pharmaceutical composition according to thepresent invention.

The term “treatment” used herein refers to all actions involved inalleviating or beneficially changing symptoms of an inflammatory diseaseby administration of the pharmaceutical composition according to thepresent invention.

In the present invention, the inflammatory disease is a disease causedby the overexpression of BLT2, and may be one or more selected fromasthma, atherosclerosis, cancer, pruritus, rheumatoid arthritis andinflammatory enteropathy, but the present invention is not limitedthereto.

Other than the diseases presented in the specification, allBLT2-associated inflammatory diseases known in the art are included asinflammatory diseases which can be prevented or treated with a compoundhaving the structure of Formula 1 of the present invention. In aparticular example, the cancer may be any cancer caused by theoverexpression of BLT2 or Ras, which is a tumor gene. The cancer may be,but is not limited to, selected from the group consisting of kidneycancer, prostatic cancer, pancreatic cancer, breast cancer, braintumors, skin cancer and liver cancer.

In the present invention, BLT2, as one among the G protein-coupledreceptor (GPCR) family, is a receptor having low affinity to LTB₄, andtherefore the composition of the present invention inhibits cell growthcaused by BLT2 to prevent or treat an inflammatory disease. Morespecifically, LTB₄-induced chemotactic motility may be inhibited byinhibiting the generation of ROS induced by BLT2 activity.

The term “inhibition” used herein refers to inhibition of a certain stepamong gene transcription, mRNA processing, translation, translocationand maturation, or inhibition of binding between proteins, activation ofa protein or signal transmission therethrough.

The pharmaceutical composition of the present invention may include apharmaceutically acceptable carrier in addition to an active ingredient.Here, the pharmaceutically acceptable carrier is conventionally used informulation, and includes, but is not limited to, lactose, dextrose,sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate,alginate, gelatin, calcium silicate, microcrystalline cellulose,polyvinyl pyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate, andmineral oil. In addition, other than the components, a lubricant, awetting agent, a sweetening agent, a flavoring agent, an emulsifier, asuspending agent or a preservative may be further included.

The pharmaceutical composition of the present invention may beadministered orally or parenterally (e.g., intravenously,subcutaneously, intraperitoneally or locally) depending on a desiredmethod, and a dose of the pharmaceutical composition may vary dependingon the condition and body weight of a patient, the severity of adisease, a drug type, an administration route and time, and may besuitably selected by one of ordinary skill in the art.

The pharmaceutical composition of the present invention is administeredat a pharmaceutically effective amount. The “pharmaceutically effectiveamount” used herein refers to an amount sufficient for treating adisease at a reasonable benefit/risk ratio applicable for medicaltreatment, and an effective dosage may be determined by parametersincluding a type of a patient's disease, severity, drug activity,sensitivity to a drug, administration time, an administration route andan excretion rate, the duration of treatment and drugs simultaneouslyused, and other parameters well known in the medical field. Thepharmaceutical composition of the present invention may be administeredseparately or in combination with other therapeutic agents, and may besequentially or simultaneously administered with a conventionaltherapeutic agent, or administered in a single or multiple dose(s). Inconsideration of all the above-mentioned parameters, it is important toachieve the maximum effect with the minimum dose without a side effect,and such a dose may be easily determined by one of ordinary skill in theart.

Specifically, the effective amount of the pharmaceutical composition ofthe present invention may be dependent on a patient's age, sex,condition and body weight, an absorption rate of the active ingredientin the body, an inactivation rate, an excretion rate, a type of disease,or a drug used in combination, and may be generally administered at0.001 to 150 mg, and preferably 0.01 to 100 mg per kg of body weightdaily or every other day, or divided into one or three dailyadministrations. However, the effective amount may vary depending on anadministration route, the severity of obesity, sex, body weight or age,and therefore, the scope of the present invention is not limited by thedose in any way.

In addition, the present invention provides a method for treating aninflammatory disease, which includes administering the pharmaceuticalcomposition to a subject. The term “subject” refers to a target diseaseto be treated, and more specifically, a mammal such as a human, or anon-human primate, a mouse, a rat, a dog, a cat, a horse and a cow.

Hereinafter, to help in understanding the present invention, exemplaryembodiments will be disclosed. However, the following examples aremerely provided to more easily understand the present invention, and thescope of the present invention is not limited to the examples.

EXAMPLES Example 1. Preparation of tert-butyl4-(4-(3-(N-phenylpentanamido)prop-1-ynyl)benzoyl)piperazine-1-carboxylate(LMT-693) Step 1: Preparation of N-phenylpentaneamide

Aniline (0.98 ml, 10.74 mmol) was dissolved in dichloromethane (20 ml),and then cooled on ice. Triethylamine (3.00 ml, 21.48 mmol) was added tothe mixture, and then stirred for 5 minutes. Valeroyl chloride (2.60 ml,21.48 mmol) was added at the same temperature, the ice was removed, andthe mixture was stirred at room temperature for 2 hours. The reactionsolution was concentrated under reduced pressure, and the residueobtained thereby was diluted with dichloromethane and washed with waterand brine. An organic solvent layer was collected, dehydrated withanhydrous magnesium sulfate (MgSO₄), filtered, and then concentratedunder reduced pressure. The concentrate was purified by silica gelcolumn chromatography (Hex:EA=10:1), thereby obtainingN-phenylpentaneamide (1.88 g, 99% yield).

Step 2: Preparation of tert-butyl4-(4-bromobenzoyl)piperazine-1-carboxylate

4-bromobenzoic acid (901 mg, 4.48 mmol) and tert-butylpiperazine-1-carboxylate (1.00 g, 5.37 mmol) were diluted inN,N-dimethylformamide (DMF; 15 ml), and stirred for 5 minutes.1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo [4,5-b]pyridinium3-oxid hexafluorophosphate (HATU; 2.04 g, 5.37 mmol) andN,N-diisopropylethylamine (DIPEA; 2.34 ml, 13.44 mmol) were added to themixture, and stirred at room temperature for 15 hours. The reactionsolution was concentrated under reduced pressure, and the residueobtained thereby was diluted with ethyl acetate and washed with waterand brine. An organic solvent layer was collected, dehydrated withanhydrous magnesium sulfate (MgSO₄), filtered, and then concentratedunder reduced pressure. The concentrate was purified by silica gelcolumn chromatography (Hex:EA=2:1), thereby obtaining tert-butyl4-(4-bromobenzoyl)piperazine-1-carboxylate (1.56 g, 94% yield).

Step 3: Preparation of tert-butyl4-(4-(3-hydroxyprop-1-ynyl)benzoyl)piperazine-1-carboxylate

The tert-butyl 4-(4-bromobenzoyl)piperazine-1-carboxylate (1.00 g, 2.71mmol) obtained in Step 2 and propargyl alcohol (0.32 ml, 5.42 mmol) weredissolved in triethylamine (12 ml), and stirred for 5 minutes.Bis(triphenylphosphine)palladium (II) dichloride (190 mg, 0.271 mmol)and copper iodide (I) (52 mg, 0.271 mmol) were added to the mixture,heated at 60° C., and refluxed to be stirred for 17 hours. The reactionmixture was cooled at room temperature, concentrated under reducedpressure, and the residue obtained thereby was diluted with ethylacetate and washed with water and brine. An organic solvent layer wascollected, dehydrated with anhydrous magnesium sulfate (MgSO₄),filtered, and then concentrated under reduced pressure. The concentratewas purified by silica gel column chromatography (Hex:EA=2:1), therebyobtaining tert-butyl4-(4-(3-hydroxyprop-1-ynyl)benzoyl)piperazine-1-carboxylate (850 mg, 91%yield).

Step 4: Preparation of tert-butyl4-(4-(3-(methylsulfonyloxy)prop-1-ynyl)benzoyl)piperazine-1-carboxylate

The tert-butyl4-(4-(3-hydroxyprop-1-ynyl)benzoyl)piperazine-1-carboxylate (600 mg,1.74 mmol) obtained in Step 3 was dissolved in dichloromethane (8 ml),and cooled on ice. Triethylamine (0.36 ml, 2.61 mmol) was added to themixture, and stirred for 5 minutes. Methanesulfonyl chloride (0.15 ml,1.92 mmol) was added at the same temperature, the ice was removed, andthe mixture was stirred at room temperature for 30 minutes. The reactionsolution was concentrated under reduced pressure, and the residueobtained thereby was diluted with dichloromethane and washed with waterand brine. An organic solvent layer was collected, dehydrated withanhydrous magnesium sulfate (MgSO₄), filtered, and then concentratedunder reduced pressure. The concentrate was purified by silica gelcolumn chromatography (Hex:EA=2:1), thereby obtaining tert-butyl4-(4-(3-(methylsulfonyloxy)prop-1-ynyl)benzoyl)piperazine-1-carboxylate(662 mg, 90% yield).

Step 5: Preparation of tert-butyl4-(4-(3-(N-phenylpentaneamido)prop-1-ynyl)benzoyl)piperazine-1-carboxylate

The N-phenylpentaneamide (185 mg, 1.04 mmol) obtained in Step 1 andsodium hydride (NaH; 75 mg, 3.12 mmol) were cooled on ice, and thentetrahydrofuran (THF; 8 ml) was added, followed by stirring for 30minutes. The tert-butyl4-(4-(3-(methylsulfonyloxy)prop-1-ynyl)benzoyl)piperazine-1-carboxylate(662 mg, 1.57 mmol) obtained in Step 4 was added to the mixture, the icewas removed, and the mixture was stirred at room temperature for 17hours. The reaction solution was concentrated under reduced pressure,and the residue obtained thereby was diluted with dichloromethane andwashed with water and brine. An organic solvent layer was collected,dehydrated with anhydrous magnesium sulfate (MgSO₄), filtered, and thenconcentrated under reduced pressure. The concentrate was purified bysilica gel column chromatography (Hex:EA=4:1), thereby obtaining a finalproduct, tert-butyl4-(4-(3-(N-phenylpentaneamido)prop-1-ynyl)benzoyl)piperazine-1-carboxylate(382 mg, 73% yield).

¹H NMR (CDCl₃, 500 MHz) δ 7.40-7.20 (9H, m), 4.65 (2H, s), 3.62-3.32(8H, br), 2.02-1.97 (2H, t), 1.52-1.48 (2H, m), 1.40 (9H, s), 1.19-1.12(2H, m), 0.76-0.72 (3H, t).

Example 2. Preparation ofN-phenyl-N-(3-(4-(piperazine-1-carbonyl)phenyl)prop-2-ynyl)pentanamide(LMT-694)

The tert-butyl4-(4-(3-(N-phenylpentaneamido)prop-1-ynyl)benzoyl)piperazine-1-carboxylate(754 mg, 1.50 mmol) obtained in Example 1 was dissolved in acetonitrile(15 ml), and stirred at room temperature for 5 minutes. Dioxane-mixedhydrochloride (4N; 3.73 ml) was added to the mixture, and stirred at thesame temperature for 1.5 hours. The reaction solution was concentratedunder reduced pressure, and a residue obtained thereby was purified bysilica gel column chromatography (CH₂Cl₂:MeOH=50:1), thereby obtaining afinal product,N-phenyl-N-(3-(4-(piperazine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide(363 mg, 60% yield).

¹H NMR (CDCl₃, 500 MHz) δ 7.48-7.30 (9H, m), 4.73 (2H, s), 3.73-3.39(4H, br), 2.97-2.86 (4H, br), 2.09-2.06 (2H, t), 1.60-1.54 (2H, m),1.25-1.19 (2H, m), 0.83-0.80 (3H, t).

Example 3. Preparation ofN-(3-(4-(4-methylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentanamide(LMT-692)

TheN-phenyl-N-(3-(4-(piperazine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide(33.3 mg, 0.072 mmol) obtained in Example 2 and potassium hydroxide(KOH; 9.09 mg, 0.108 mmol) were dissolved in N,N-dimethylformamide (DMF;1 ml), and stirred at room temperature for 5 minutes. Iodomethane (9 μl,0.144 mmol) was added to the mixture, and stirred at the sametemperature for 17 hours. The reaction solution was concentrated underreduced pressure, and a residue obtained thereby was diluted withdichloromethane and washed with water and brine. An organic solventlayer was collected, dehydrated with anhydrous magnesium sulfate(MgSO₄), filtered, and then concentrated under reduced pressure. Theconcentrate was purified by silica gel column chromatography(CH₂Cl₂:MeOH=20:1), thereby obtaining a final product,N-(3-(4-(4-methylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentaneamide(6 mg, 20% yield).

¹H NMR (CDCl₃, 400 MHz) δ 7.40-7.21 (9H, m), 4.65 (2H, s), 3.71-3.34(4H, br), 2.41-2.25 (4H, br), 2.25 (3H, s), 2.02-1.99 (2H, t), 1.54-1.46(2H, m), 1.18-1.12 (2H, m), 0.76-0.71 (3H, t).

Example 4. Preparation ofN-(3-(4-(4-ethylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentaneamide(LMT-695)

TheN-phenyl-N-(3-(4-(piperazine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide(24.8 mg, 0.061 mmol) obtained in Example 3 and potassium hydroxide(8.62 mg, 0.154 mmol) were dissolved in N,N-dimethylformamide (DMF; 1ml), and stirred at room temperature for 5 minutes. Iodoethane (20 al,0.246 mmol) was added to the mixture, and stirred at the sametemperature for 17 hours. The reaction solution was concentrated underreduced pressure, and a residue obtained thereby was diluted withdichloromethane and washed with water and brine. An organic solventlayer was collected, dehydrated with anhydrous magnesium sulfate(MgSO₄), filtered, and then concentrated under reduced pressure. Theconcentrate was purified by silica gel column chromatography(CH₂Cl₂:MeOH=20:1), thereby obtaining a final product,N-(3-(4-(4-ethylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentaneamide(17.9 mg, 68% yield).

¹H NMR (CDCl₃, 400 MHz) δ 7.40-7.20 (9H, m), 4.65 (2H, s), 3.73-3.35(4H, br), 2.44-2.31 (6H, m), 2.03-1.99 (2H, t), 1.54-1.46 (2H, m),1.20-1.13 (2H, m), 1.05-1.01 (3H, t), 0.78-0.73 (3H, t).

Example 5. Preparation ofN-(3-(4-(4-isopropylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentaneamide(LMT-696)

TheN-phenyl-N-(3-(4-(piperazine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide(106 mg, 0.263 mmol) obtained in Example 2 and sodium bicarbonate (27mg, 0.316 mmol) were cooled on ice, and then N,N-dimethylformamide (DMF;2 ml) was added, followed by stirring for 1 hour. 2-iodopropane (30 al,0.316 mmol) was added to the mixture, the ice was removed, and then themixture was heated at 60° C., refluxed and stirred for 24 hours. Thereaction solution was cooled at room temperature and concentrated underreduced pressure, and a residue obtained thereby was diluted withethylacetate and washed with water and brine. An organic solvent layerwas collected, dehydrated with anhydrous magnesium sulfate (MgSO₄),filtered, and then concentrated under reduced pressure. The concentratewas purified by silica gel column chromatography(Hex:EA:MeOH:TEA=12:12:1:0.1), thereby obtaining a final product,N-(3-(4-(4-isopropylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentaneamide(66.8 mg, 57% yield).

¹H NMR (CDCl₃, 500 MHz) δ 7.47-7.30 (9H, m), 4.73 (2H, s), 3.78-3.40(4H, br), 2.75-2.72 (1H, m), 2.59-2.44 (4H, br), 2.09-2.06 (2H, t),1.59-1.56 (2H, m), 1.25-1.20 (2H, m), 1.06-1.04 (6H, d), 0.83-0.80 (3H,t).

Example 6. Preparation ofN-(3-(4-(4-(2-hydroxyethyl)piperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentaneamide(LMT-827)

TheN-phenyl-N-(3-(4-(piperazine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide(56 mg, 0.139 mmol) obtained in Example 2 and potassium carbonate (77mg, 0.556 mmol) were dissolved in acetonitrile (3 ml), and stirred atroom temperature for 5 minutes. 2-bromoethanol (99 al, 1.39 mmol) wasadded to the mixture, the ice was removed, and then the mixture washeated at 60° C., refluxed and stirred for 17 hours. An organic solventlayer was collected, dehydrated with anhydrous magnesium sulfate(MgSO₄), filtered, and then concentrated under reduced pressure. Theconcentrate was purified by silica gel column chromatography(CH₂Cl₂:MeOH=50:1), thereby obtaining a final product,N-(3-(4-(4-(2-hydroxyethyl)piperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentaneamide(52 mg, 84% yield).

¹H NMR (CDCl₃, 500 MHz) δ 7.48-7.30 (9H, m), 4.73 (2H, s), 3.79 (2H,br), 3.66-3.64 (2H, t), 3.43 (2H, br), 2.60-2.46 (7H, br), 2.10-2.07(2H, t), 1.59-1.56 (2H, m), 1.25-1.22 (2H, m), 0.83-0.80 (3H, t).

Example 7. Preparation ofN-(3-(4-(4-(cyclopropylmethyl)piperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentaneamide(LMT-828)

TheN-phenyl-N-(3-(4-(piperazine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide(50 mg, 0.124 mmol) obtained in Example 2 and potassium carbonate (51mg, 0.372 mmol) were dissolved in N,N-dimethylformamide (DMF; 2 ml), andstirred at room temperature for 5 minutes. Cyclopropylmethyl bromide (15al, 0.145 mmol) was added to the mixture, heated at 80° C., refluxed andstirred for 4 hours. An organic solvent layer was collected, dehydratedwith anhydrous magnesium sulfate (MgSO₄), filtered, and thenconcentrated under reduced pressure. The concentrate was purified bysilica gel column chromatography (CH₂Cl₂:MeOH=50:1), thereby obtaining afinal product,N-(3-(4-(4-(cyclopropylmethyl)piperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentanamide(16 mg, 28% yield).

¹H NMR (CDCl₃, 500 MHz) δ 7.48-7.28 (9H, m), 4.73 (2H, s), 3.82-3.45(4H, br), 2.63-2.49 (4H, br), 2.32-2.31 (2H, d), 2.09-2.06 (2H, t),1.60-1.56 (2H, m), 1.25-1.20 (3H, m), 0.83-0.80 (3H, t), 0.55-0.53 (2H,m), 0.12-0.11 (2H, m).

Example 8. Preparation ofN-(3-(4-(4-cyclohexylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentaneamide(LMT-830)

A final product,N-(3-(4-(4-cyclohexylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentaneamide,was obtained (20 mg, 33% yield) by the same method described in Example7 using theN-phenyl-N-(3-(4-(piperazine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide(50 mg, 0.124 mmol) obtained in Example 2 and iodocyclohexane (19 al,0.145 mmol).

¹H NMR (CDCl₃, 500 MHz) δ 7.48-7.31 (9H, m), 4.73 (2H, s), 3.77-3.39(4H, br), 2.63-2.49 (4H, br), 2.31-2.28 (1H, m), 2.09-2.06 (2H, m),1.91-1.79 (4H, m), 1.65-1.54 (3H, m), 1.28-1.16 (6H, m), 1.13-1.08 (1H,m), 0.83-0.80 (3H, t).

Example 9. Preparation ofN-(3-(4-(4-(cyclohexylmethyl)piperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentanamide(LMT-831)

A final product,N-(3-(4-(4-(cyclohexylmethyl)piperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentaneamide,was obtained (35 mg, 56% yield) by the same method as described inExample 7 using theN-phenyl-N-(3-(4-(piperazine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide(50 mg, 0.124 mmol) obtained in Example 2 and bromomethylcyclohexane (20al, 0.145 mmol).

¹H NMR (CDCl₃, 500 MHz) δ 7.48-7.28 (9H, m), 4.73 (2H, s), 3.76-3.38(4H, br), 2.45-2.31 (4H, br), 2.15-2.13 (2H, m), 2.09-2.06 (2H, m),1.77-1.66 (5H, m), 1.59-1.56 (2H, m), 1.47-1.45 (1H, m), 1.25-1.17 (5H,m), 0.90-0.80 (5H, t).

Example 10. Preparation ofN-(3-(4-(4-isobutylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentanamide(LMT-832)

A final product,N-(3-(4-(4-isobutylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentaneamide,was obtained (34 mg, 60% yield) by the method as described in Example 7using theN-phenyl-N-(3-(4-(piperazine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide(50 mg, 0.124 mmol) obtained in Example 2 and 1-iodo-2-methylpropane (17al, 0.145 mmol).

¹H NMR (CDCl₃, 500 MHz) δ 7.47-7.28 (9H, m), 4.73 (2H, s), 3.76-3.39(4H, br), 2.46-2.32 (4H, br), 2.11-2.07 (4H, m), 1.79-1.76 (1H, m),1.59-1.56 (2H, m), 1.25-1.20 (2H, m), 0.91-0.89 (6H, d), 0.83-0.80 (3H,t).

Example 11. Preparation ofN-phenyl-N-(3-(4-(4-(prop-2-ynyl)piperazine-1-carbonyl)phenyl)prop-2-ynyl)pentanamide(LMT-833)

TheN-phenyl-N-(3-(4-(piperazine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide(50 mg, 0.124 mmol) obtained in Example 2 and potassium carbonate (51mg, 0.372 mmol) were dissolved in N,N-dimethylformamide (DMF; 2 ml), andstirred at room temperature for 5 minutes. Propargyl bromide (12 al,0.145 mmol) was added to the mixture, and stirred at room temperaturefor 17 hours. The reaction solution was filtered to remove a solid andconcentrated under reduced pressure, and the concentrate was purified bysilica gel column chromatography (CH₂Cl₂:MeOH=100:1), thereby obtaininga final product,N-phenyl-N-(3-(4-(4-(prop-2-ynyl)piperazine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide(28 mg, 51% yield).

¹H NMR (CDCl₃, 500 MHz) δ 7.48-7.28 (9H, m), 4.73 (2H, s), 3.82-3.44(4H, br), 3.36 (2H, s), 2.65-2.51 (4H, br), 2.30 (1H, s), 2.10-2.07 (2H,m), 1.60-1.54 (2H, m), 1.26-1.19 (2H, m), 0.83-0.80 (3H, t).

Example 12. Preparation ofN-(3-(4-(4-cyanopiperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentanamide(LMT-829)

TheN-phenyl-N-(3-(4-(piperazine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide(50 mg, 0.124 mmol) obtained in Example 2 and trimethylsilyl cyanide (49al, 0.372 mmol) were dissolved in acetonitrile (2 ml), and stirred atroom temperature for 5 minutes. Sodium hypochloride (43 al, 0.620 mmol)was added to the mixture, heated at 80° C., refluxed, and stirred for 12hours. The reaction solution was cooled at room temperature, filtered toremove a solid, and concentrated under reduced pressure. The obtainedresidue was diluted with ethyl acetate, and washed with water and brine.An organic solvent layer was collected, dehydrated with anhydrousmagnesium sulfate (MgSO₄), filtered, and then concentrated under reducedpressure. The concentrate was purified by silica gel columnchromatography (CH₂Cl₂:MeOH=200:1), thereby obtaining a final product,N-(3-(4-(4-cyanopiperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentanamide(11 mg, 21% yield).

¹H NMR (CDCl₃, 500 MHz) δ 7.48-7.30 (9H, m), 4.73 (2H, s), 3.81-3.26(8H, br), 2.09-2.06 (2H, t), 1.60-1.54 (2H, m), 1.25-1.19 (2H, m),0.83-0.80 (3H, t).

Example 13. Preparation of tert-butyl4-(4-(3-(N-(3-fluorophenyl)pentanamido)prop-1-ynyl)benzoyl)piperazine-1-carboxylate(LMT-884) Step 1: Preparation of N-(3-fluorophenyl)pentanamide

N-(3-fluorophenyl)pentaneamide was obtained (1.74 g, 99% yield) using3-fluoroaniline (0.87 ml, 9.00 mmol) and valeroyl chloride (2.18 ml,18.00 mmol).

Step 2: Preparation of tert-butyl4-(4-(3-(N-(3-fluorophenyl)pentaneamido)prop-1-ynyl)benzoyl)piperazine-1-carboxylate

A final product, tert-butyl4-(4-(3-(N-(3-fluorophenyl)pentaneamido)prop-1-ynyl)benzoyl)piperazine-1-carboxylate,was obtained (197 mg, 73% yield) by the same method as described in Step5 of Example 1 using the N-(3-fluorophenyl)pentaneamide (101 mg, 0.519mmol) obtained in Step 1 and the tert-butyl4-(4-(3-(methylsulfonyloxy)prop-1-ynyl)benzoyl)piperazine-1-carboxylate(329 mg, 0.779 mmol) obtained in Step 4 of Example 1.

¹H NMR (CDCl₃, 400 MHz) δ 7.44-7.32 (5H, m), 7.12-7.06 (3H, m), 4.72(2H, s), 3.73-3.38 (8H, br), 2.10-2.07 (2H, t), 1.59-1.57 (2H, m), 1.47(9H, s), 1.23-1.20 (2H, m), 0.83-0.80 (3H, t).

Example 14. Preparation ofN-(3-fluorophenyl)-N-(3-(4-(piperazine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide(LMT-885)

A final product,N-(3-fluorophenyl)-N-(3-(4-(piperazine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide,was obtained (94 mg, 58% yield) by the same method as described inExample 12 using the tert-butyl4-(4-(3-(N-(3-fluorophenyl)pentaneamido)prop-1-ynyl)benzoyl)piperazine-1-carboxylate(200 mg, 0.383 mmol) obtained in Example 13.

¹H NMR (CDCl₃, 400 MHz) δ 7.45-7.32 (5H, m), 7.15-7.07 (3H, m), 4.72(2H, s), 3.75-3.37 (4H, br), 2.94-2.80 (4H, br), 2.10-2.07 (2H, t), 1.89(1H, br), 1.60-1.57 (2H, m), 1.25-1.24 (2H, m), 0.84-0.82 (3H, t).

Example 15. Preparation ofN-(3-fluorophenyl)-N-(3-(4-(4-isopropylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide(LMT-886)

A final product,N-(3-fluorophenyl)-N-(3-(4-(4-isopropylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide,was obtained (20 mg, 40% yield) using theN-(3-fluorophenyl)-N-(3-(4-(piperazine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide(46 mg, 0.109 mmol) obtained in Example 14 and 2-iodopropane (0.375 ml,3.77 mmol).

¹H NMR (CDCl₃, 400 MHz) δ 7.44-7.32 (5H, m), 7.15-7.06 (3H, m), 4.72(2H, s), 3.78-3.41 (4H, br), 2.75-2.72 (1H, m), 2.59-2.45 (4H, br),2.11-2.09 (2H, t), 1.60-1.57 (2H, m), 1.25-1.22 (2H, m), 1.06-1.05 (6H,d), 0.85-0.82 (3H, t).

Example 16. Preparation of tert-butyl4-(4-(3-(N-(4-fluorophenyl)pentaneamido)prop-1-ynyl)benzoyl)piperazine-1-carboxylate(LMT-839) Step 1: Preparation of N-(4-fluorophenyl)pentaneamide

N-(4-fluorophenyl)pentaneamide was obtained (174 mg, 99% yield) by thesame method as described in Step 1 of Example 1 using 4-fluoroaniline(85 μl, 0.90 mmol) and valeroyl chloride (0.22 ml, 1.80 mmol).

Step 2: Preparation of tert-butyl4-(4-(3-(N-(4-fluorophenyl)pentanamido)prop-1-ynyl)benzoyl)piperazine-1-carboxylate

A final product, tert-butyl4-(4-(3-(N-(4-fluorophenyl)pentaneamido)prop-1-ynyl)benzoyl)piperazine-1-carboxylate,was obtained (2.28 g, 73% yield) by the same method as described in Step5 of Example 1 using the N-(4-fluorophenyl)pentaneamide (1.17 g, 6.00mmol) obtained in Step 1 and the tert-butyl4-(4-(3-(methylsulfonyloxy)prop-1-ynyl)benzoyl)piperazine-1-carboxylate(3.80 g, 8.99 mmol) obtained in Step 4 of Example 1.

¹H NMR (CDCl₃, 400 MHz) δ 7.39-7.29 (6H, m), 7.17-7.14 (2H, m), 4.71(2H, s), 3.73-3.38 (8H, br), 2.07-2.04 (2H, t), 1.60-1.54 (2H, m), 1.47(9H, s), 1.25-1.19 (2H, m), 0.84-0.81 (3H, t).

Example 17. Preparation ofN-(4-fluorophenyl)-N-(3-(4-(piperazine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide(LMT-840)

A final product,N-(4-fluorophenyl)-N-(3-(4-(piperazine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide,was obtained (1.06 g, 58% yield) by the same method as described inExample 12 using the tert-butyl4-(4-(3-(N-(4-fluorophenyl)pentaneamido)prop-1-ynyl)benzoyl)piperazine-1-carboxylate(2.28 g, 4.37 mmol) obtained in Example 16.

¹H NMR (CDCl₃, 400 MHz) δ 7.38-7.30 (6H, m), 7.17-7.15 (2H, m), 4.71(2H, s), 3.77-3.40 (4H, br), 2.96-2.79 (5H, br), 2.06-2.03 (2H, t),1.57-1.54 (2H, m), 1.25-1.22 (2H, m), 0.84-0.82 (3H, t).

Example 18. Preparation ofN-(4-fluorophenyl)-N-(3-(4-(4-isopropylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide(LMT-841)

A final product,N-(4-fluorophenyl)-N-(3-(4-(4-isopropylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide,was obtained (465 mg, 40% yield) by the same method as described inExample 15 using theN-(4-fluorophenyl)-N-(3-(4-(piperazine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide(1.06 g, 2.51 mmol) obtained in Example 17 and 2-iodopropane (0.375 ml,3.77 mmol).

¹H NMR (CDCl₃, 400 MHz) δ 7.37-7.28 (6H, m), 7.17-7.13 (2H, m), 4.71(2H, s), 3.78-3.40 (4H, br), 2.75-2.72 (1H, m), 2.59-2.45 (4H, br),2.07-2.04 (2H, t), 1.58-1.55 (2H, m), 1.25-1.21 (2H, m), 1.06-1.04 (6H,d), 0.84-0.81 (3H, t).

Example 19. Preparation ofN-(3-(4-(morpholine-4-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentaneamide(LMT-682) Step 1: (4-bromophenyl)(morpholino)methanone

(4-bromophenyl)(morpholino)methanone was obtained (455 mg, yield 99%) bythe same method as described in Step 2 of Example 1 using 4-bromobenzoicacid (340 mg, 1.70 mmol) and morpholine (0.18 ml, 2.04 mmol).

Step 2: Preparation of(4-(3-hydroxyprop-1-ynyl)phenyl)(morpholino)methanone

(4-(3-hydroxyprop-1-ynyl)phenyl)(morpholino)methanone was obtained (371mg, yield 90%) by the same method as described in Step 3 of Example 1using the (4-bromophenyl)(morpholino)methanone (455 mg, 1.68 mmol)obtained in Step 1 and propargyl alcohol (0.196 ml, 3.36 mmol).

Step 3: Preparation of 3-(4-(morpholine-4-carbonyl)phenyl)prop-2-ynylmethanesulfonate

3-(4-(morpholine-4-carbonyl)phenyl)prop-2-ynyl methanesulfonate wasobtained (391 mg, yield 80%) by the same method as described in Step 4of Example 1 using the(4-(3-hydroxyprop-1-ynyl)phenyl)(morpholino)methanone (371 mg, 1.51mmol) obtained in Step 2 and methanesulfonyl chloride (0.128 ml, 1.66mmol).

Step 4: Preparation ofN-(3-(4-(morpholine-4-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentaneamide

A final product,N-(3-(4-(morpholine-4-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentaneamide,was obtained (371 mg, yield 90%) by the same method as described in Step5 of Example 1 using the N-phenylpentaneamide (143 mg, 0.81 mmol)obtained in Step 1 of Example 1 and the3-(4-(morpholine-4-carbonyl)phenyl)prop-2-ynyl methanesulfonate (391 mg,1.21 mmol) obtained in Step 3.

¹H NMR (400 MHz, CDCl₃) δ 7.48-7.28 (m, 9H), 4.73 (s, 2H), 3.74-3.66(br, 6H), 3.43 (br, 2H), 2.08 (m, 2H), 1.57 (m, 2H), 1.22 (m, 2H), 0.81(t, 3H).

Example 20. Preparation ofN-phenyl-N-(3-(4-(piperidine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide(LMT-683) Step 1: Preparation of(4-bromophenyl)(piperidin-1-yl)methanone

(4-bromophenyl)(piperidin-1-yl)methanone was obtained (458.2 mg, 100%yield) by the same method as described in Step 2 of Example 1 using4-bromobenzoic acid (318 mg, 1.58 mmol) and piperidine (0.21 ml, 1.90mmol).

Step 2: Preparation of(4-(3-hydroxyprop-1-ynyl)phenyl)(piperidin-1-yl)methanone

(4-(3-hydroxyprop-1-ynyl)phenyl)(piperidin-1-yl)methanone was obtained(374 mg, yield 90%) by the same method as described in Step 3 of Example1 using the (4-bromophenyl)(piperidine-1-yl)methanone (458.2 mg, 1.71mmol) obtained in Step 1 and propargyl alcohol (0.199 ml, 3.42 mmol).

Step 3: Preparation of 3-(4-(piperidine-1-carbonyl)phenyl)prop-2-ynylmethanesulfonate

3-(4-(piperidine-1-carbonyl)phenyl)prop-2-ynyl methanesulfonate wasobtained (396 mg, yield 80%) by the same method as described in Step 4of Example 1 using the(4-(3-hydroxyprop-1-ynyl)phenyl)pyridine-1-yl)methanone (374 mg, 1.54mmol) obtained in Step 2 and methanesulfonyl chloride (0.131 ml, 1.69mmol).

Step 4: Preparation ofN-phenyl-N-(3-(4-(piperidine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide

A final product,N-phenyl-N-(3-(4-(piperidine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide,was obtained (50 mg, 15% yield) by the same method as described in Step5 of Example 1 using the N-phenylpentaneamide (146 mg, 0.82 mmol)obtained in Step 1 of Example 1 and the3-(4-(pyridine-1-carbonyl)phenyl)prop-2-ynyl methanesulfonate (396 mg,1.23 mmol) obtained in Step 3.

¹H NMR (CDCl₃, 400 MHz) δ 7.40-7.13 (9H, m), 4.65 (2H, s, CH₂),3.62-3.24 (4H, br), 2.02-1.99 (2H, t), 1.60 (4H, br), 1.52-1.46 (2H, m),1.44 (2H, br), 1.20-1.08 (2H, m), 0.78-0.73 (3H, t).

Example 21. Preparation ofN,N-diethyl-4-(3-(N-phenylpentaneamido)prop-1-ynyl)benzamide (LMT-883)Step 1: Preparation of 4-bromo-N,N-diethylbenzamide

4-bromo-N,N-diethylbenzamide was obtained (700 mg, 69% yield) by thesame method as described in Step 2 of Example 1 using 4-bromobenzoicacid (800 mg, 3.98 mmol) and diethylamine (0.49 ml, 4.78 mmol).

Step 2: Preparation of N,N-diethyl-4-(3-hydroxyprop-1-ynyl)benzamide

N,N-diethyl-4-(3-hydroxyprop-1-ynyl)benzamide was obtained (425 mg, 67%yield) by the same method as described in Step 3 of Example 1 using the4-bromo-N,N-diethylbenzamide (700 mg, 2.73 mmol) obtained in Step 1 andpropargyl alcohol (0.32 ml, 5.47 mmol).

Step 3: Preparation of 3-(4-(diethylcarbamoyl)phenyl)prop-2-ynylmethanesulfonate

3-(4-(diethylcarbamoyl)phenyl)prop-2-ynyl methanesulfonate was obtained(483 mg, 85% yield) by the same method as described in Step 4 of Example1 using the N,N-diethyl-4-(3-hydroxyprop-1-ynyl)benzamide (425 mg, 1.84mmol) obtained in Step 2 and methanesulfonyl chloride (0.16 ml, 2.02mmol).

Step 4: Preparation ofN,N-diethyl-4-(3-(N-phenylpentanamido)prop-1-ynyl)benzamide

A final product,N,N-diethyl-4-(3-(N-phenylpentaneamido)prop-1-ynyl)benzamide, wasobtained (81 mg, 39% yield) by the same method as described in Step 5 ofExample 1 using the N-phenylpentaneamide (94 mg, 0.530 mmol) obtained inStep 1 of Example 1 and the 3-(4-(diethylcarbamoyl)phenyl)prop-2-ynylmethanesulfonate (246 mg, 0.795 mmol) obtained in Step 3.

¹H NMR (CDCl₃, 400 MHz) δ 7.48-7.30 (9H, m), 4.73 (2H, s), 3.53-3.23(4H, br), 2.10-2.07 (2H, t), 1.59-1.57 (2H, m), 1.23-1.10 (8H, m),0.83-0.80 (3H, t).

Example 22. Preparation ofN-phenyl-N-(3-(3-(piperazine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide(LMT-837) Step 1: Preparation of tert-butyl4-(3-bromobenzoyl)piperazine-1-carboxylate

Tert-butyl 4-(3-bromobenzoyl)piperazine-1-carboxylate was obtained (2.75g, 99% yield) by the same method described in Step 2 of Example 1 using3-bromobenzoic acid (1.50 g, 7.46 mmol) and tert-butylpiperazine-1-carboxylate (1.67 g, 8.95 mmol).

Step 2: Preparation of tert-butyl4-(3-(3-hydroxyprop-1-ynyl)benzoyl)piperazine-1-carboxylate

Tert-butyl 4-(3-(3-hydroxyprop-1-ynyl)benzoyl)piperazine-1-carboxylatewas obtained (2.32 g, 90% yield) by the same method as described in Step3 of Example 1 using the tert-butyl4-(3-bromobenzoyl)piperazine-1-carboxylate (2.75 g, 7.45 mmol) obtainedin Step 1 and propargyl alcohol (0.87 ml, 14.89 mmol).

Step 3: Preparation of tert-butyl4-(3-(3-(methylsulfonyloxy)prop-1-ynyl)benzoyl)piperazine-1-carboxylate

Tert-butyl4-(3-(3-(methylsulfonyloxy)prop-1-ynyl)benzoyl)piperazine-1-carboxylatewas obtained (1.78 g, 63% yield) by the same method as described in Step4 of Example 1 using the tert-butyl4-(3-(3-hydroxyprop-1-ynyl)benzoyl)piperazine-1-carboxylate (2.32 g,6.75 mmol) obtained in Step 2 and methanesulfonyl chloride (0.58 ml,7.42 mmol).

Step 4: Preparation of tert-butyl4-(3-(3-(N-phenylpentaneamido)prop-1-ynyl)benzoyl)piperazine-1-carboxylate

A final product tert-butyl4-(3-(3-(N-phenylpentaneamido)prop-1-ynyl)benzoyl)piperazine-1-carboxylatewas obtained (323 mg, 70% yield) by the same method as described in Step5 of Example 1 using the N-phenylpentaneamide (275 mg, 1.55 mmol)obtained in Step 1 of Example 1 and the tert-butyl4-(3-(3-(methylsulfonyloxy)prop-1-ynyl)benzoyl)piperazine-1-carboxylate(982 mg, 2.33 mmol) obtained in Step 3.

Step 5: Preparation ofN-phenyl-N-(3-(3-(piperazine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide(LMT-837)

A final product,N-phenyl-N-(3-(3-(piperazine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide,was obtained (152 mg, 58% yield) by the same method as described inExample 2 using the tert-butyl4-(3-(3-(N-phenylpentaneamido)prop-1-ynyl)benzoyl)piperazine-1-carboxylate(323 mg, 0.62 mmol) obtained in Step 4.

¹H NMR (CDCl₃, 400 MHz) δ 7.45-7.26 (9H, m), 4.69 (2H, s), 3.75-3.36(4H, br), 2.94-2.80 (4H, br), 2.59 (1H, br), 2.07-2.04 (2H, t),1.56-1.53 (2H, m), 1.22-1.18 (2H, m), 0.80-0.77 (3H, t).

Example 23. Preparation ofN-(3-(3-(4-methylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentaneamide(LMT-838)

TheN-phenyl-N-(3-(3-(piperazine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide(50 mg, 0.124 mmol) obtained in Example 22 and formaldehyde (37% in H₂O;1.5 ml) were dissolved in formic acid (2.0 ml), heated at 100° C.,refluxed, and stirred for 4 hours. The reaction solution wasconcentrated under reduced pressure, and titrated by adding a sodiumhydroxide aqueous solution (2.0 M). Afterward, the reaction product wasdiluted with dichloromethane, and washed with water and brine. Anorganic solvent layer was collected, dehydrated with anhydrous magnesiumsulfate (MgSO₄), filtered, and then concentrated under reduced pressure.The concentrate was purified by silica gel column chromatography(CH₂Cl₂:MeOH=20:1), thereby obtainingN-(3-(3-(4-methylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentaneamide(24 mg, 46% yield).

¹H NMR (CDCl₃, 400 MHz) δ 7.46-7.29 (9H, m), 4.70 (2H, s), 3.79-3.39(4H, br), 2.48-2.32 (7H, br), 2.08-2.05 (2H, t), 1.57-1.54 (2H, m),1.23-1.19 (2H, m), 0.81-0.78 (3H, t).

Example 24. Preparation ofN-(3-(3-(4-isopropylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentaneamide(LMT-842)

A final product,N-(3-(3-(4-isopropylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentaneamide,was obtained (67 mg, 40% yield) by the same method as described inExample 5 using theN-phenyl-N-(3-(3-(piperazine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide(152 mg, 0.36 mmol) obtained in Example 22 and 2-iodopropane (90 al,0.90 mmol).

¹H NMR (CDCl₃, 400 MHz) δ 7.47-7.29 (9H, m), 4.71 (2H, s), 3.79-3.40(4H, br), 2.78-2.75 (1H, m), 2.60-2.46 (4H, br), 2.09-2.06 (2H, t),1.58-1.55 (2H, m), 1.24-1.20 (2H, m), 1.07-1.05 (6H, d), 0.82-0.79 (3H,t).

Example 25. Preparation of tert-butyl4-(3-(3-(N-(4-fluorophenyl)pentaneamido)prop-1-ynyl)benzoyl)piperazine-1-carboxylate(LMT-887)

A final product,tert-butyl-4-(3-(3-(N-(4-fluorophenyl)pentaneamido)prop-1-ynyl)benzoyl)piperazine-1-carboxylate,was obtained (625 mg, 85% yield) by the same method as described in Step5 of Example 1 using the N-(4-fluorophenyl)pentaneamide (275 mg, 1.41mmol) obtained in Step 1 of Example 16 and the tert-butyl4-(3-(3-(methylsulfonyloxy)prop-1-ynyl)benzoyl)piperazine-1-carboxylate(894 mg, 2.12 mmol) obtained in Step 3 of Example 22.

¹H NMR (CDCl₃, 400 MHz) δ 7.33-7.20 (6H, m), 7.09-7.06 (2H, m), 4.62(2H, s), 3.66-3.31 (8H, br), 2.00-1.97 (2H, t), 1.52-1.49 (2H, m), 1.47(9H, s), 1.18-1.13 (2H, m), 0.76-0.73 (3H, t).

Example 26. Preparation ofN-(4-fluorophenyl)-N-(3-(3-(piperazine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide(LMT-888)

A final product,N-(4-fluorophenyl)-N-(3-(3-(piperazine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide,was obtained (356 mg, 58% yield) by the same method as described inExample 2 using thetert-butyl-4-(3-(3-(N-(4-fluorophenyl)pentaneamido)prop-1-ynyl)benzoyl)piperazine-1-carboxylate(762 mg, 1.46 mmol) obtained in Example 25.

¹H NMR (CDCl₃, 400 MHz) δ 7.39-7.29 (6H, m), 7.17-7.14 (2H, m), 4.70(2H, s), 3.77-3.39 (4H, br), 2.98-2.85 (4H, br), 2.08-2.05 (2H, t),1.60-1.54 (2H, m), 1.25-1.19 (2H, m), 0.84-0.81 (3H, t).

Example 27. Preparation ofN-(4-fluorophenyl)-N-(3-(3-(4-isopropylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide(LMT-889)

A final product,N-(4-fluorophenyl)-N-(3-(3-(4-isopropylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide,was obtained (66 mg, 40% yield) by the same method as described inExample 5 using theN-(4-fluorophenyl)-N-(3-(3-(piperazine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide(150 mg, 0.356 mmol) obtained in Example 26 and 2-iodopropane (53 al,0.534 mmol).

¹H NMR (CDCl₃, 400 MHz) δ 7.37-7.29 (6H, m), 7.16-7.13 (2H, m), 4.70(2H, s), 3.79-3.39 (4H, br), 2.75-2.73 (1H, m), 2.59-2.45 (4H, br),2.07-2.04 (2H, t), 1.58-1.55 (2H, m), 1.25-1.21 (2H, m), 1.06-1.05 (6H,d), 0.84-0.81 (3H, t).

Example 28. Preparation ofN-(3-(4-hydroxyphenyl)prop-2-ynyl)-N-phenylpentaneamide (LMT-890) Step1: Preparation of N-(prop-2-ynyl)aniline

Aniline (2.94 ml, 32.21 mmol) and potassium carbonate (4.90 g, 35.43mmol) were dissolved in acetonitrile (40 ml), and stirred for 5 minutes.Propargyl bromide (3.05 ml, 35.43 mmol) was added to the mixture, andstirred at room temperature for 17 hours. The reaction solution wasfiltered to remove a solid, and concentrated under reduced pressure. Theconcentrate was purified by silica gel column chromatography (Hex),thereby obtaining N-(prop-2-ynyl)aniline (2.23 g, 53% yield).

Step 2: Preparation of N-phenyl-N-(prop-2-ynyl)pentaneamide

N-phenyl-N-(prop-2-ynyl)pentaneamide was obtained (1.29 g, 95% yield) bythe same method as described in Step 1 of Example 1 using theN-(prop-2-ynyl)aniline (828 mg, 6.31 mmol) obtained in Step 1 andvaleroyl chloride (1.53 ml, 12.62 mmol).

Step 3: Preparation ofN-(3-(4-hydroxyphenyl)prop-2-ynyl)-N-phenylpentaneamide

The N-phenyl-N-(prop-2-ynyl)pentaneamide (550 mg, 2.55 mmol) obtained inStep 2 and 4-iodophenol (422 mg, 1.92 mmol) were dissolved intriethylamine (15 ml), and stirred for 5 minutes.Bis(triphenylphosphine)palladium (II) dichloride (89 mg, 0128 mmol) andcopper iodide (I) (49 mg, 0.255 mmol) were added to the mixture, heatedat 50° C., refluxed, and stirred for 5 hours. The reaction solution wascooled at room temperature and concentrated under reduced pressure, andthe obtained residue was diluted with ethyl acetate and washed withwater and brine. An organic solvent layer was collected, dehydrated withanhydrous magnesium sulfate (MgSO₄), filtered, and then concentratedunder reduced pressure. The concentrate was purified by silica gelcolumn chromatography (Hex:EA=10:1), thereby obtaining a final product,N-(3-(4-hydroxyphenyl)prop-2-ynyl)-N-phenylpentaneamide (590 mg, 75%yield).

¹H NMR (CDCl₃, 400 MHz) δ 8.15 (1H, br), 7.47-6.83 (9H, m), 4.66 (2H,s), 2.13-2.10 (2H, t), 1.60-1.54 (2H, m), 1.22-1.17 (2H, m), 0.80-0.77(3H, t).

Example 29. Preparation of2-(4-(3-(N-phenylpentaneamido)prop-1-ynyl)phenoxy)acetic acid (LMT-891)Step 1: Preparation of ethyl(2-(4-(3-(N-phenylpentaneamido)prop-1-ynyl)phenoxy)acetate

The N-(3-(4-hydroxyphenyl)prop-2-ynyl)-N-phenylpentaneamide (590 mg,1.92 mmol) obtained in Example 28 and potassium carbonate (796 mg, 5.76mmol) were dissolved in acetonitrile (15 ml), and stirred for 30minutes. Ethyl bromoacetate (0.23 ml, 2.11 mmol) was added to themixture, and stirred at room temperature for 17 hours. The reactionsolution was filtered to remove a solid and concentrated under reducedpressure, and the obtained residue was diluted with ethyl acetate andwashed with water and brine. An organic solvent layer was collected,dehydrated with anhydrous magnesium sulfate (MgSO₄), filtered, and thenconcentrated under reduced pressure. The concentrate was purified bysilica gel column chromatography (Hex:EA=10:1), thereby obtaining ethyl2-(4-(3-(N-phenylpentaneamido)prop-1-ynyl)phenoxy)acetate (530 mg, 70%yield).

Step 2: Preparation of2-(4-(3-(N-phenylpentaneamido)prop-1-ynyl)phenoxy)acetic acid

The ethyl 2-(4-(3-(N-phenylpentaneamido)prop-1-ynyl)phenoxy)acetate (530mg, 1.35 mmol) obtained in Step 1 was dissolved in ethanol (15 ml), andstirred at room temperature for 5 minutes. 2N sodium hydroxide (NaOH;0.50 ml) was added to the mixture, heated at 80° C., refluxed andstirred for 3 hours. The reaction solution was cooled at roomtemperature and concentrated under reduced pressure, and the obtainedresidue was diluted with ethyl acetate and washed with water and brine.An organic solvent layer was collected, dehydrated with anhydrousmagnesium sulfate (MgSO₄), filtered, and then concentrated under reducedpressure. The concentrate was purified by silica gel columnchromatography (CH₂Cl₂:MeOH=100:1), thereby obtaining a final product,2-(4-(3-(N-phenylpentaneamido)prop-1-ynyl)phenoxy)acetic acid (246 mg,50% yield).

¹H NMR (CDCl₃, 400 MHz) δ 7.47-6.78 (9H, m), 4.68 (2H, s), 4.58 (2H, s),2.11-2.08 (2H, t), 1.58-1.53 (2H, m), 1.23-1.18 (2H, m), 0.81-0.78 (3H,t).

Example 30. Preparation of tert-butyl4-(5-(3-(N-phenylpentanamido)prop-1-yn-1-yl)picolinoyl)piperazine-1-carboxylate(LMT-834)

A final product, tert-butyl4-(5-(3-(N-phenylpentaneamido)prop-1-yn-1-yl)picolinoyl)piperazine-1-carboxylate,was obtained (36 mg, 35% yield) using the same method as described inStep 3 of Example 28 using the N-phenyl-N-(prop-2-ynyl)pentaneamideobtained in Step 2 of Example 28 and tert-butyl4-(5-bromopicolinoyl)piperazine-1-carboxylate (86 mg, 0.4 mmol).

¹H NMR (400 MHz, CDCl₃) δ 8.43 (s, 1H), 7.64 (dd, 1H), 7.54 (dd, 1H),7.36 (dd, 3H), 7.18 (m, 2H), 4.68 (s, 2H), 3.69 (br, 2H), 3.53-3.38 (br,6H), 2.01 (m, 2H), 1.52 (m, 2H), 1.39 (s, 9H), 1.18 (m, 2H), 0.73 (t,3H).

Example 31. Preparation ofN-phenyl-N-(3-(6-(piperazine-1-carbonyl)pyridine-3-yl)prop-2-yn-1-yl)pentaneamide(LMT-835)

The tert-butyl4-(5-(3-((N-phenylpentaneamido)prop-1-yn-1-yl)picolinoyl)piperazine-1-carboxylate(35 mg, 0.69 mmol) obtained in Example 30 was dissolved in acetonitrile(15 ml), and stirred at room temperature for 5 minutes. Dioxane-mixedhydrochloride (4N; 3.73 ml) was added to the mixture, and stirred at thesame temperature for 1.5 hours. The reaction solution was concentratedunder reduced pressure, and the obtained residue was purified by silicagel column chromatography (CH₂Cl₂:MeOH=50:1), thereby obtaining a finalproduct,N-phenyl-N-(3-(6-(piperazine-1-carbonyl)pyridine-3-yl)prop-2-yn-1-yl)pentaneamide(18 mg, 64% yield).

¹H NMR (400 MHz, CDCl₃) δ 8.43 (s, 1H), 7.66 (dd, 1H), 7.49 (dd, 1H),7.36 (dd, 3H), 7.21 (m, 2H), 4.67 (s, 2H), 3.70 (br, 2H), 3.48 (br, 2H),2.90 (br, 2H), 2.81 (br, 2H), 2.01 (m, 2H), 1.49 (m, 2H), 1.17 (m, 2H),0.73 (t, 3H).

Example 32. Preparation ofN-(3-(6-(4-isopropylpiperazine-1-carbonyl)pyridine-3-yl)prop-2-yn-1-yl)-N-phenylpentaneamide(LMT-836)

TheN-phenyl-N-(3-(6-(piperazine-1-carbonyl)pyridine-3-yl)prop-2-yn-1-yl)pentaneamide(57.7 mg, 0.14 mmol) obtained in Example 31 and sodium bicarbonate (27mg, 0.316 mmol) were cooled on ice, and N,N-dimethylformamide (DMF; 2ml) was added, followed by stirring for 1 hour. 2-iodopropane (30 al,0.316 mmol) was added to the mixture, the ice was removed, and theresulting product was heated at 60° C., refluxed, and stirred for 24hours. The reaction solution was cooled at room temperature andconcentrated under reduced pressure, and the obtained residue wasdiluted with ethyl acetate and washed with water and brine. An organicsolvent layer was collected, dehydrated with anhydrous magnesium sulfate(MgSO₄), filtered, and then concentrated under reduced pressure. Theconcentrate was purified by silica gel column chromatography(Hex:EA:MeOH:TEA=12:12:1:0.1), thereby obtaining a final product,N-(3-(6-isopropylpiperazine-1-carbonyl)pyridine-3-yl)prop-2-yn-1-yl)-N-phenylpentaneamide(32.3 mg, 51% yield).

¹H NMR (400 MHz, CDCl₃) δ 8.44 (s, 1H), 7.65 (dd, 1H), 7.51 (dd, 1H),7.39 (dd, 3H), 7.22 (m, 2H), 4.68 (s, 2H), 3.74 (br, 2H), 3.52 (br, 2H),2.67 (m, 1H), 2.55 (br, 2H), 2.41 (br, 2H), 2.02 (m, 2H), 1.50 (m, 2H),1.15 (m, 2H), 0.98 (d, 6H), 0.74 (t, 3H).

Example 33. Preparation ofN,N-diethyl-4-(3-(N-(3-fluorophenyl)pentaneamido)prop-1-yn-1-yl)benzamide(LMT-926) Step 1: Preparation of 4-bromo-N,N-diethylbenzamide

4-Bromobenzoic acid (5.00 g, 24.9 mmol) was dissolved inN,N-dimethylformamide (100.00 ml), and mixed with diisopropylamine (13ml, 74.6 mmol). 1-Hydroxybenzotriazole hydrate (7.15 mg, 37.30 mmol) and1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (5.04 mg,37.30 mmol) were added to the mixture, and stirred for 5 minutes.Diethylamine (3.1 ml, 37.3 mmol) was added to the mixture, and stirredat room temperature for 12 hours. The reaction solution was concentratedunder reduced pressure, and the obtained residue was diluted withdichloromethane and washed with water and brine. An organic solventlayer was collected, dehydrated with anhydrous magnesium sulfate(MgSO₄), filtered, and then concentrated under reduced pressure. Theconcentrate was purified by silica gel column chromatography(Hex:EA=3:1), thereby obtaining 4-bromo-N,N-diethylbenzamide (5.70 g,89% yield).

Step 2: Preparation of N,N-diethyl-4-(3-hydroxyprop-1-ynyl)benzamide

The 4-bromo-N,N-diethylbenzamide (5.70 mg, 22.3 mmol) obtained in Step 1and propargyl alcohol (2.60 ml, 44.5 mmol) were dissolved intriethylamine (100.00 ml), and stirred for 5 minutes.Bis(triphenylphosphine)palladium (II) dichloride (1.60 mg, 2.23 mmol)and copper iodide (I) (1.60 mg, 2.23 mmol) were added to the mixture,heated at 60° C., refluxed, and stirred for 17 hours. The reactionsolution was cooled at room temperature, concentrated under reducedpressure, and the obtained residue was diluted with ethyl acetate andwashed with water and brine. An organic solvent layer was collected,dehydrated with anhydrous magnesium sulfate (MgSO₄), filtered, and thenconcentrated under reduced pressure. The concentrate was purified bysilica gel column chromatography (Hex:EA=1:1), thereby obtainingN,N-diethyl-4-(3-hydroxyprop-1-ynyl)benzamide (5.16 mg, 99.9% yield).

Step 3: Preparation of 3-(4-(diethylcarbamoyl)phenyl)prop-2-yn-1-ylmethanesulfonate

N,N-diethyl-4-(3-hydroxyprop-1-ynyl)benzamide (5.16 mg, 22.3 mmol) wasdissolved in dichloromethane (100 ml), and cooled on ice. Triethylamine(4.80 ml, 34.4 mmol) was added to the mixture, and stirred for 5minutes. Methanesulfonyl chloride (1.95 ml, 25.2 mmol) was added at thesame temperature, the ice was removed, and the mixture was stirred atroom temperature for 30 minutes. The reaction solution was concentratedunder reduced pressure, and the obtained residue was diluted with ethylacetate and washed with water and brine. An organic solvent layer wascollected, dehydrated with anhydrous magnesium sulfate (MgSO₄),filtered, and then concentrated under reduced pressure. The concentratewas purified by silica gel column chromatography (Hex:EA=2:1), therebyobtaining 3-(4-(diethylcarbamoyl)phenyl)prop-2-yn-1-yl methanesulfonate(4.2 mg, 59% yield).

Step 4: Preparation ofN,N-diethyl-4-(3-(N-(3-fluorophenyl)pentaneamido)prop-1-yn-1-yl)benzamide

N-(3-fluorophenyl)pentaneamide (200 mg, 1.02 mmol) was dissolved intetrahydrofuran (10.00 ml), and cooled on ice. Sodium hydroxide (73 mg,3.06 mmol) was added to the mixture, and stirred for 1 hour. The3-(4-(N,N-diethylcarbamoyl)phenyl)prop-2-ynyl methanesulfonate (475 mg,1.54 mmol) obtained in Step 3 was added at the same temperature, and themixture was stirred at room temperature for 4 hours. The reactionsolution was concentrated under reduced pressure, and the obtainedresidue was diluted with dichloromethane and washed with water andbrine. An organic solvent layer was collected, dehydrated with anhydrousmagnesium sulfate (MgSO₄), filtered, and then concentrated under reducedpressure. The concentrate was purified by silica gel columnchromatography (Hex:EA=2:1), thereby obtainingN,N-diethyl-4-(3-(N-(3-fluorophenyl)pentaneamido)prop-1-yn-1-yl)benzamide(291.7 mg, 70% yield).

1H NMR (CDCl₃, 500 MHz) δ 7.43 (1H, t, J=7.5 Hz and 15.0 Hz, aromatic),7.33 (4H, m, aromatic), 7.09 (3H, m, aromatic), 4.71 (2H, s, CH₂), 3.53(2H, s, CH₂), 3.23 (2H, s, CH₂), 2.10 (2H, m, CH₂), 1.59 (2H, m, CH₂),1.24 (2H, m, CH₂), 1.10 (6H, m, (CH₃)₂), 0.83 (3H, t, J=7.5 Hz and 15.0Hz, CH₃).

Example 34. Preparation ofN,N-diethyl-4-(3-(N-(4-fluorophenyl)pentaneamido)prop-1-yn-1-yl)benzamide(LMT-927)

N,N-diethyl-4-(3-(N-(4-fluorophenyl)pentaneamido)prop-1-yn-1-yl)benzamidewas obtained (291.7 mg, 70% yield) by the same method as described inStep 4 of Example 33 using N-(4-fluorophenyl)pentaneamide (200 mg, 1.02mmol) and 3-(4-N,N-(diethylcarbamoyl)phenyl)prop-2-yn-1-ylmethanesulfonate.

¹H NMR (CDCl₃, 500 MHz) δ 7.31 (2H, d, J=3.5 Hz, aromatic), 7.29 (4H, d,J=3.0 Hz, aromatic), 7.15 (2H, t, J=8.5 Hz and 17.0 Hz, aromatic), 4.71(2H, s, CH₂), 3.53 (2H, s, CH₂), 3.23 (2H, s, CH₂), 2.06 (2H, t, J=7.5Hz and 15.0 Hz, CH₂), 1.57 (2H, m, CH₂), 1.22 (2H, m, CH₂), 1.10 (6H, s,(CH₃)₂), 0.82 (3H, t, J=7.5 Hz and 15.0 Hz, CH₃).

Example 35. Preparation ofN-(3-(4-(N,N-diethylsulfamoyl)phenyl)prop-2-ynyl)-N-phenylpentaneamide(LMT-946) Step 1: Preparation of 4-bromo-N,N-diethylbenzenesulfonamide

4-Bromobenzenesulfonyl chloride (1.00 g, 3.91 mmol) was dissolved indichloromethane (30.00 ml), and cooled on ice. Diethylamine (1.19 ml,11.54 mmol) was added to the mixture and stirred for 5 minutes, the icewas removed, and then the mixture was stirred at room temperature for 12hours. The reaction solution was concentrated under reduced pressure,and the obtained residue was diluted with dichloromethane and washedwith water and brine. An organic solvent layer was collected, dehydratedwith anhydrous magnesium sulfate (MgSO₄), filtered, and thenconcentrated under reduced pressure. The concentrate was purified bysilica gel column chromatography (Hex:EA=3:1), thereby obtaining4-bromo-N,N-diethylbenzenesulfonamide (1.10 g, 96% yield).

¹H NMR (CDCl₃, 500 MHz) δ 7.67 (4H, m), 3.23 (4H, q, J=7.0 Hz), 1.13(6H, t).

Step 2: Preparation ofN,N-diethyl-4-(3-hydroxyprop-1-ynyl)benzenesulfonamide

The 4-bromo-N,N-diethylbenzenesulfonamide (500.00 mg, 1.71 mmol)obtained in Step 1 and propargyl alcohol (0.20 ml, 3.42 mmol) weredissolved in triethylamine (10.00 ml), and stirred for 5 minutes.Bis(triphenylphosphine)palladium (II) dichloride (119.32 mg, 0.17 mmol)and copper iodide (I)(32.37 mg, 0.17 mmol) were added to the mixture,heated at 60° C., refluxed, and stirred for 17 hours. The reactionsolution was cooled at room temperature and concentrated under reducedpressure, and the obtained residue was diluted with ethyl acetate andwashed with water and brine. An organic solvent layer was collected,dehydrated with anhydrous magnesium sulfate (MgSO₄), filtered, and thenconcentrated under reduced pressure. The concentrate was purified bysilica gel column chromatography (Hex:EA=1:1), thereby obtainingN,N-diethyl-4-(3-hydroxyprop-1-ynyl)benzenesulfonamide (286.00 mg, 63%yield).

¹H NMR (CDCl₃, 500 MHz) δ 7.66 (2H, d, J=8.0 Hz), 7.42 (2H, d, J=8.0Hz), 4.44 (2H, s), 3.16 (4H, q, J=7.0 Hz), 1.05 (6H, t).

Step 3: Preparation of 3-(4-(N,N-diethylsulfamoyl)phenyl)prop-2-ynylmethanesulfonate

The N,N-diethyl-4-(3-hydroxyprop-1-ynyl)benzenesulfonamide (273.00 mg,1.02 mmol) obtained in Step 2 was dissolved in dichloromethane (10 ml),and cooled on ice. Triethylamine (0.21 ml, 1.53 mmol) was added to themixture, and stirred for 5 minutes. Methanesulfonyl chloride (0.09 ml,1.12 mmol) was added at the same temperature, the ice was removed, andthe mixture was stirred at room temperature for 30 minutes. The reactionsolution was concentrated under reduced pressure, and the obtainedresidue was diluted with ethyl acetate and washed with water and brine.An organic solvent layer was collected, dehydrated with anhydrousmagnesium sulfate (MgSO₄), filtered, and then concentrated under reducedpressure. The concentrate was purified by silica gel columnchromatography (Hex:EA=2:1), thereby obtaining3-(4-(N,N-diethylsulfamoyl)phenyl)prop-2-ynyl methanesulfonate (285.00mg, 80% yield).

¹H NMR (CDCl₃, 500 MHz) δ 7.73 (2H, d, J=8.5 Hz), 7.53 (2H, d, J=8.5Hz), 5.05 (2H, s), 3.19 (4H, q, J=7.0 Hz), 3.12 (3H, s), 1.07 (6H, t).

Step 4: Preparation of4-(3-bromoprop-1-ynyl)-N,N-diethylbenzenesulfonamide

The 3-(4-(N,N-diethylsulfamoyl)phenyl)prop-2-ynyl methanesulfonate(260.00 mg, 0.75 mmol) obtained in Step 3 was dissolved intetrahydrofuran (20.00 ml), and cooled on ice. Lithium bromide (196.28mg, 2.26 mmol) was added to the mixture at the same temperature, andstirred for 4 hours. The reaction solution was concentrated underreduced pressure, and the obtained residue was diluted with ethylacetate and washed with water and brine. An organic solvent layer wascollected, dehydrated with anhydrous magnesium sulfate (MgSO₄),filtered, and then concentrated under reduced pressure. The concentratewas purified using a filter, thereby obtaining4-(3-bromoprop-1-ynyl)-N,N-diethylbenzenesulfonamide (240.00 mg, 97%).

Step 5: Preparation ofN,N-diethyl-4-(3-(phenylamino)prop-1-ynyl)benzenesulfonamide

The 4-(3-bromoprop-1-ynyl)-N,N-diethylbenzenesulfonamide (248.00 mg,0.75 mmol) obtained in Step 4 and potassium carbonate (93.98 mg, 0.68mmol) were dissolved in acetonitrile (15.00 ml), and stirred for 30minutes. Aniline (0.06 ml, 0.68 mmol) was added to the mixture, andstirred at room temperature for 9 hours. The reaction solution wasconcentrated under reduced pressure, and the obtained residue wasdiluted with ethyl acetate and washed with water and brine. An organicsolvent layer was collected, dehydrated with anhydrous magnesium sulfate(MgSO₄), filtered, and then concentrated under reduced pressure. Theconcentrate was purified by silica gel column chromatography(Hex:EA=2:1), thereby obtainingN,N-diethyl-4-(3-(phenylamino)prop-1-ynyl)benzenesulfonamide (190.00 mg,81% yield).

¹H NMR (CDCl₃, 500 MHz) δ 7.71 (2H, d, J=8.5 Hz), 7.46 (2H, d, J=8.5Hz), 7.23 (2H, t), 6.80 (1H, t), 6.73 (2H, d, J=7.5 Hz), 4.15 (2H, s),3.21 (4H, m), 1.10 (6H, t).

Step 6: Preparation ofN-(3-(4-(N,N-diethylsulfamoyl)phenyl)prop-2-ynyl)-N-phenylpenteamide

The N,N-diethyl-4-(3-(phenylamino)prop-1-ynyl)benzenesulfonamide (174.00mg, 0.51 mmol) obtained in Step 5 was dissolved in dichloromethane(15.00 ml), and cooled on ice. Triethylamine (0.14 ml, 1.02 mmol) wasadded to the mixture, and stirred for 5 minutes. Valeroyl chloride (0.06ml, 0.53 mmol) was added at the same temperature, the ice was removed,and the mixture was stirred at room temperature for 4 hours. Thereaction solution was concentrated under reduced pressure, and theobtained residue was diluted with ethyl acetate and washed with waterand brine. An organic solvent layer was collected, dehydrated withanhydrous magnesium sulfate (MgSO₄), filtered, and then concentratedunder reduced pressure. The concentrate was purified by silica gelcolumn chromatography (Hex:EA=2:1), thereby obtainingN-(3-(4-(N,N-diethylsulfamoyl)phenyl)prop-2-ynyl)-N-phenylpentaneamide(153.00 mg, 70% yield).

¹H NMR (CDCl₃, 500 MHz) δ 7.74 (2H, d, J=8.0 Hz), 7.51 (4H, m), 7.43(3H, m), 4.73 (2H, s), 3.21 (4H, s), 2.11 (2H, t), 1.52 (2H, m), 1.21(2H, m), 0.10 (6H, t), 0.80 (3H, t).

Example 36. Preparation ofN-(3-(4-(N-isopropylsulfamoyl)phenyl)prop-2-ynyl)-N-phenylpentaneamide(LMT-947) Step 1: Preparation of 4-bromo-N-isopropylbenzenesulfonamide

4-Bromobenzenesulfonyl chloride (1.00 g, 3.91 mmol) was dissolved indichloromethane (10.00 ml), and isopropylamine (0.40 ml, 4.69 mmol) andpyridine (0.41 ml, 5.09 mmol) were added to the mixture and stirred atroom temperature for 2 hours. The reaction solution was concentratedunder reduced pressure, and the obtained residue was diluted with ethylacetate and washed with water and brine. An organic solvent layer wascollected, dehydrated with anhydrous magnesium sulfate (MgSO₄),filtered, and then concentrated under reduced pressure. The concentratewas purified by silica gel column chromatography (Hex:EA=3:1), therebyobtaining 4-bromo-N-isopropylbenzenesulfonamide (730.00 mg, 67% yield).

¹H NMR (CDCl₃, 500 MHz) δ 7.76-7.64 (4H, m), 3.47 (1H, m), 1.09 (6H, d,J=7.0 Hz).

Step 2: Preparation of4-(3-hydroxyprop-1-ynyl)-N-isopropylbenzenesulfonamide

4-(3-hydroxyprop-1-ynyl)-N-isopropylbenzenesulfonamide was obtained(420.00 mg, 92% yield) by the same method as described in Step 2 ofExample 35 using the 4-bromo-N-isopropylbenzenesulfonamide (500.00 mg,1.80 mmol) obtained in Step 1 and propargyl alcohol (0.21 ml, 3.59mmol).

¹H NMR (CDCl₃, 500 MHz) δ 7.81 (2H, d, J=8.0 Hz), 7.49 (2H, d, J=8.5Hz), 4.51 (2H, s), 3.44 (1H, m), 1.06 (6H, d, J=6.5 Hz).

Step 3: Preparation of 3-(4-(N-isopropylsulfamoyl)phenyl)prop-2-ynylmethanesulfonate

3-(4-(N-isopropylsulfamoyl)phenyl)prop-2-ynyl methanesulfonate wasobtained (330.00 mg, 61% yield) by the same method as described in Step3 of Example 35 using the4-(3-hydroxyprop-1-ynyl)-N-isopropylbenzenesulfonamide (410.00 mg, 1.62mmol) obtained in Step 2 and methanesulfonyl chloride (0.14 ml, 1.78mmol).

¹H NMR (CDCl₃, 500 MHz) δ 7.86 (2H, d, J=8.5 Hz), 7.59 (2H, d, J=8.5Hz), 5.10 (2H, s), 3.47 (1H, m) 3.17 (3H, s), 1.08 (6H, d, J=6.5 Hz).

Step 4: Preparation of4-(3-bromoprop-1-ynyl)-N-isopropylbenzenesulfonamide

4-(3-bromoprop-1-ynyl)-N-isopropylbenzenesulfonamide was obtained(190.00 mg, 95% yield) by the same method as described in Step 4 ofExample 35 using the 3-(4-(N-isopropylsulfamoyl)phenyl)prop-2-ynylmethanesulfonate (210.00 mg, 0.63 mmol) obtained in Step 3 and lithiumbromide (165.00 mg, 1.90 mmol).

Step 5: Preparation ofN-isopropyl-4-(3-(phenylamino)prop-1-ynyl)benzenesulfonamide

N-isopropyl-4-(3-(phenylamino)prop-1-ynyl)benzenesulfonamide wasobtained (157 mg, 83% yield) using the4-(3-bromoprop-1-ynyl)-N-isopropylbenzenesulfonamide (199.00 mg, 0.63mmol) obtained in Step 4 and aniline (0.05 ml, 0.57 mmol).

¹H NMR (CDCl₃, 500 MHz) δ 7.80 (2H, d, J=9.0 Hz), 7.50 (2H, d, J=8.5Hz), 7.25 (2H, m), 6.82 (1H, t), 6.76 (2H, d, J=8.0 Hz), 4.64 (1H, d,J=8.0 Hz), 4.19 (2H, s), 3.45 (1H, m), 1.06 (6H, d, J=6.0 Hz).

Step 6: Preparation ofN-(3-(4-(N-isopropylsulfamoyl)phenyl)prop-2-ynyl)-N-phenylpentaneamide

N-(3-(4-(N-isopropylsulfamoyl)phenyl)prop-2-ynyl)-N-phenylpentaneamidewas obtained (94.00 mg, 49.5% yield) by the same method as described inStep 6 of Example 35 using theN-isopropyl-4-(3-(phenylamino)prop-1-ynyl)benzenesulfonamide (150.00 mg,0.46 mmol) obtained in Step 2 and valeroyl chloride (0.06 ml, 0.48mmol).

¹H NMR (CDCl₃, 500 MHz) δ 7.78 (2H, d, J=8.5 Hz), 7.51-7.37 (7H, m),4.72 (2H, s), 3.33 (1H, m), 2.10 (2H, t), 1.51 (2H, m), 1.19 (2H, m),0.99 (6H, d, J=6.5 Hz), 0.78 (3H, t)

Example 37. Preparation of tert-butyl4-(3-(N-phenylpentaneamido)prop-1-yn-1-yl)benzoate (LMT-1012) Step 1:Preparation of N-phenylpentaneamide

Aniline (10.00 ml, 107.40 mmol) was dissolved in dichloromethane (150ml), and cooled on ice. Triethylamine (30.00 ml, 214.80 mmol) was addedto the mixture, and stirred for 5 minutes. Valeroyl chloride (16.00 ml,128.90 mmol) was added at the same temperature, the ice was removed, andthe mixture was stirred at room temperature for 2 hours. The reactionsolution was concentrated under reduced pressure, and the obtainedresidue was diluted with ethyl acetate and washed with water and brine.An organic solvent layer was collected, dehydrated with anhydrousmagnesium sulfate (MgSO₀₄), filtered, and then concentrated underreduced pressure. The concentrate was purified by silica gel columnchromatography (Hex:EA=10:1), thereby obtaining N-phenylpentaneamide(19.1 g, 99.9% yield).

¹H NMR (CDCl₃, 500 MHz) δ 7.55 (2H, d, J=8.0 Hz, aromatic), 7.29 (2H, t,J=7.5 Hz and 15.0 Hz, aromatic), 7.09 (1H, t, J=7.0 Hz and 14.5 Hz,aromatic), 2.35 (2H, t, J=8.0 Hz and 15.5 Hz, CH₂), 1.70 (2H, m, CH₂),1.37 (2H, m, CH₂), 0.92 (3H, t, J=7.0 Hz and 14.5 Hz, CH₃).

Step 2: Preparation of N-phenyl-N-(prop-2-yn-1-yl)pentaneamide

The N-phenylpentaneamide (19.10 g, 107.40 mmol) obtained in Step 1 wasdissolved in N,N-dimethylformamide (DMF; 100 ml), and a reaction systemwas substituted with nitrogen, sodium hydride (5.20 g, 214.80 mmol) wasadded at a sub-zero temperature and then stirred for 2 hours. Propargylbromide (18.10 ml, 214.80 mmol) was added to the mixture, and stirred ata sub-zero temperature for 2 hours. Water was added to the reactionsolution at a sub-zero temperature, which was then diluted with ethylacetate, and washed with water and brine. An organic solvent layer wascollected, dehydrated with anhydrous magnesium sulfate (MgSO₄),filtered, and then concentrated under reduced pressure. The concentratewas purified by silica gel column chromatography (Hex:EA=9:1), therebyobtaining N-phenyl-N-(prop-2-yn-1-yl)pentanamide (19.20 g, 83% yield).

¹H NMR (CDCl₃, 500 MHz) δ 7.42 (2H, m, aromatic), 7.37 (1H, d, J=7.0 Hz,aromatic), 7.26 (2H, m, aromatic), 4.47 (2H, d, J=2.0 Hz, CH₂), 2.03(2H, t, J=7.0 Hz and 15.5 Hz, CH₂), 1.53 (2H, m, CH₂), 1.20 (2H, m,CH₂), 0.77 (3H, t, J=7.5 Hz and 15 Hz, CH₃).

Step 3: Preparation of tert-butyl 4-iodobenzoate

Thionyl chloride (2.30 ml, 32.30 mmol) and N,N-dimethylformamide (DMF)(0.02 ml, 0.20 mmol) were added to 4-iodobenzoic acid (1.00 g, 4.00mmol), and then the reaction system was substituted with nitrogen,heated to 75° C., refluxed, and then stirred for 1 hour. The reactionsolution was concentrated under reduced pressure, the obtained residuewas dissolved in tetrahydrofuran (5 ml), and then a potassiumtert-butoxide 1M solution in THF (4.5 ml) was slowly added at a sub-zerotemperature, and stirred for 30 minutes. The reaction solution wasconcentrated under reduced pressure, and the obtained residue wasdiluted with ethyl acetate and washed with water and brine. An organicsolvent layer was collected, dehydrated with anhydrous magnesium sulfate(MgSO₄), filtered, and then concentrated under reduced pressure. Theconcentrate was purified by silica gel column chromatography(Hex:EA=9:1), thereby obtaining 2-(trimethylsilyl)ethyl 4-iodobenzoate(14.00 g, 99.9% yield).

¹H NMR (CDCl₃, 400 MHz) δ 7.77 (2H, d, J=7.5 Hz, aromatic), 7.69 (2H, d,J=8.0 Hz, aromatic), 1.59 (9H, s, (CH₃)₃).

Step 4: Preparation of tert-butyl4-(3-(N-phenylpentaneamido)prop-1-yn-1-yl)benzoate

The N-phenyl-N-(pro-2-yn-1-yl)pentaneamide (2.30 g, 10.70 mmol) obtainedin Step 2 was added to a solution in which the tert-butyl 4-iodobenzoate(4.90 g, 16.00 mmol) obtained in Step 3 was dissolved in tetrahydrofuran(30 ml), and a reaction system was substituted with nitrogen, followedby stirring at room temperature for 5 minutes. Triethylamine (24 ml),bis(triphenylphosphine)palladium (II) dichloride (75.00 mg, 0.10 mmol)and copper iodide (I) (41.00 mg, 0.21 mmol) were added to the mixture,and stirred at room temperature for 16 hours. The resulting product wasconcentrated under reduced pressure, and the obtained residue wasdiluted with dichloromethane and washed with water and brine. An organicsolvent layer was collected, dehydrated with anhydrous magnesium sulfate(MgSO₄), filtered, and then concentrated under reduced pressure. Theconcentrate was purified by silica gel column chromatography(Hex:EA=4:1), thereby obtaining tert-butyl4-(3-(N-phenylpentaneamido)prop-1-yn-1-yl)benzoate (3.30 g, 78.3%yield).

¹H-NMR (500 MHz, CDCl₃): δ 7.88 (2H, d, J=8.0 Hz, aromatic), 7.45 (2H,m, aromatic), 7.39 (1H, d, J=7.0 Hz, aromatic), 7.35 (2H, d, J=8.0 Hz,aromatic), 7.30 (2H, d, J=5.0 Hz, aromatic), 4.72 (2H, s, CH₂), 2.07(2H, t, J=7.5 Hz and 15 Hz, CH₂), 1.56 (2H, m, CH₂), 1.51 (9H, s,(CH₃)₃), 1.22 (2H, m, CH₂), 0.80 (3H, t, 7.5 Hz and 15 Hz7.5 Hz and 15Hz, CH₃).

Example 38. Preparation of4-(3-(N-phenylpentaneamido)prop-1-yn-1-yl)benzoic acid (LMT-1013)

The tert-butyl 4-(3-(N-phenylpentaneamido)prop-1-yn-1-yl)benzoate (2.00g, 5.10 mmol) obtained in Example 37 was dissolved in acetonitrile (48ml), and stirred at a sub-zero temperature for 5 minutes.Trifluoroacetic acid (12 ml) was slowly added to the solution, andstirred at room temperature for 48 hours. The reaction solution wasconcentrated under reduced pressure, and the obtained residue wasdiluted with ethyl acetate and washed with water and brine. An organicsolvent layer was collected, dehydrated with anhydrous magnesium sulfate(MgSO₄), filtered, and then concentrated under reduced pressure. Theconcentrate was purified by silica gel column chromatography(HEX:EA=2:1), thereby obtaining4-(3-(N-phenylpentaneamido)pro-1-yn-1-yl)benzoic acid (1.6 g, 95%yield).

¹H-NMR (500 MHz, MeOD): δ 7.95 (2H, d, J=8.5 Hz, aromatic), 7.51 (2H, m,aromatic), 7.45 (1H, m, aromatic), 7.44 (2H, d, J=8.5 Hz, aromatic),7.39 (2H, d, J=8.0 Hz, aromatic), 4.73 (2H, s, CH₂), 2.11 (2H, t, CH₃),1.21 (2H, m, CH₂), 0.83 (3H, t, CH₃).

Example 39. Preparation ofN-ethyl-4-(3-(N-phenylpentaneamido)prop-1-yn-1-yl)benzamide (LMT-1017)

The 4-(3-(N-phenylpentaneamido)prop-1-yn-1-yl)benzoic acid (80.00 mg,0.24 mmol) obtained in Example 38 was dissolved in N,N-dimethylformamide(DMF; 0.70 ml), ethylaminehydrochloride (29.20 mg, 0.36 mmol) and1-hydroxybenzotriazole hydrate (48.4 mg, 0.36 mmol), which were stirredin triethylamine (0.70 ml) for 1 hour, and1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (55.5 mg,0.36 mmol) were added to the solution and then stirred at roomtemperature for 16 hours. The reaction solution was diluted with ethylacetate and washed with water and brine. An organic solvent layer wascollected, dehydrated with anhydrous magnesium sulfate (MgSO₄),filtered, and then concentrated under reduced pressure. The concentratewas purified by silica gel column chromatography (Hex:EA=1:1), therebyobtaining N-ethyl-4-(3-(N-phenylpentaneamido)prop-1-yn-1-yl)benzamide(56.8 mg, 66% yield).

¹H NMR (CDCl₃, 500 MHz) δ 7.69 (2H, d, J=8.5 Hz, aromatic), 7.39 (7H, m,aromatic), 4.72 (2H, s, CH₂), 3.49 (2H, t, J=6.0 Hz and 13.0 Hz, CH₂),2.08 (2H, t, J=7.5 Hz and 15.0 Hz, CH₂), 1.57 (2H, m, CH₂), 1.23 (2H, m,CH₂), 0.81 (3H, t, J=7.5 Hz and 14.5 Hz, CH₃).

Example 40. Preparation ofN-(2-(dimethylamino)ethyl)-4-(3-(N-phenylpentaneamido)prop-1-yn-1-yl)benzamide(LMT-1016)

N-(2-(diethylamino)ethyl)-4-(3-(N-phenylpentaneamido)prop-1-yn-1-yl)benzamidewas obtained (71.72 mg, 74% yield) by the same method as described inExample 39 using the 4-(3-(N-phenylpentaneamido)pro-1-yn-1-yl)benzoicacid (80.00 mg, 0.24 mmol) obtained in Example 38 andN,N-dimethylethane-1,2-diamine (0.04 ml, 0.36 mmol).

¹H NMR (CDCl₃, 500 MHz) δ 7.71 (2H, d, J=8.0 Hz, aromatic), 7.38 (7H, m,aromatic), 4.70 (2H, s, CH₂), 3.48 (2H, m, CH₂), 2.50 (2H, t, J=6.0 Hzand 11.5 Hz, CH₂), 2.24 (6H, s, (CH₃)₂), 2.05 (2H, t, J=7.5 Hz and 15.0Hz, CH₂), 1.54 (2H, m, CH₂), 1.20 (2H, m, CH₂), 0.78 (3H, t, J=7.0 Hzand 14.0 Hz, CH₃).

Example 41. Preparation of ethyl2-(4-(3-(N-phenylpentaneamido)prop-1-yn-1-yl)benzamido)acetate(LMT-1014)

Ethyl 2-(4-(3-(N-phenylpentaneamido)prop-1-yn-1-yl)benzamido)acetate wasobtained (102.18 mg, 54% yield) by the same method as described inExample 39 using the 4-(3-(N-phenylpentaneamido)pro-1-yn-1-yl)benzoicacid (150.00 mg, 0.45 mmol) obtained in Example 38 and glycineethylesterhydrochloride (93.70 mg, 0.67 mmol).

¹H NMR (CDCl₃, 500 MHz) δ 7.74 (2H, d, J=8.0 Hz, aromatic), 7.37 (7H, m,aromatic), 4.22 (4H, m, (CH₂)₂), 2.07 (2H, t, J=7.5 Hz and 15.0 Hz,CH₂), 1.56 (2H, m, CH₂), 1.30 (5H, m, CH₃, CH₂), 1.22 (2H, m, CH₂), 0.80(3H, t, J=7.5 Hz and 14.5 Hz, CH₃).

Example 42. Preparation of2-(4-(3-(N-phenylpentaneamido)prop-1-yn-1-yl)benzamido)acetic acid(LMT-1015)

The ethyl 2-(4-(3-(N-phenylpentaneamido)prop-1-yn-1-yl)benzamido)acetate(46.20 mg, 0.10 mmol) obtained in Example 41 and a 2M sodium hydroxideaqueous solution (0.07 ml, 0.14 mmol) were dissolved in methanol (0.1ml), and stirred at room temperature for 30 minutes. The acidity of thereaction solution was increased using hydrochloric acid, which was thendiluted with ethylacetate, and washed with water and brine. An organicsolvent layer was collected, dehydrated with anhydrous magnesium sulfate(MgSO₄), filtered, and then concentrated under reduced pressure. Theconcentrate was recrystallized using hexene and ethylacetate, therebyobtaining 2-(4-(3-(N-phenylpentaneamido)prop-1-yn-1-yl)benzamido)aceticacid (20.80 mg, 53% yield).

¹H NMR (CDCl₃, 500 MHz) δ 7.71 (2H, d, J=8.5 Hz, aromatic), 7.40 (7H, m,aromatic), 4.71 (2H, s, CH₂), 4.23 (2H, d, J=5.0 Hz, CH₂), 2.10 (2H, t,J=7.5 Hz and 15.5 Hz, CH₂), 1.56 (2H, m, CH₂), 1.22 (2H, m, CH₂), 0.80(3H, t, J=7.0 Hz and 14.5 Hz, CH₃).

Example 43. Preparation of methyl2-(4-(3-(N-phenylpentaneamido)prop-1-yn-1-yl)benzamido)propanoate(LMT-1018)

Methyl 2-(4-(3-(N-phenylpentaneamido)prop-1-yn-1-yl)benzamido)propanoatewas obtained (110.00 mg, 42% yield) using the4-(3-(N-phenylpentaneamido)pro-1-yn-1-yl)benzoic acid (100.00 mg, 0.30mmol) obtained in Example 38 and L-alaninemethylester hydrochloride(83.70 mg, 0.60 mmol).

¹H NMR (CDCl₃, 500 MHz) δ 7.72 (2H, d, J=8.5 Hz, aromatic), 7.37 (7H, m,aromatic), 4.76 (1H, t, J=7.5 Hz and 14.5 Hz CH), 4.71 (2H, s, CH₂),3.76 (3H, s, CH₃), 2.06 (2H, t, J=7.5 Hz and 15.5 Hz, CH₂), 1.53 (2H, m,CH₂), 1.50 (3H, d, J=7.5 Hz, CH₃), 1.20 (2H, m, CH₂), 0.79 (3H, t, J=7.5Hz and 15.0 Hz, CH₃).

Example 44. Preparation of2-(4-(3-(N-phenylpentaneamido)prop-1-yn-1-yl)benzamido)propanoic acid(LMT-1019)

2-(4-(3-(N-phenylpentaneamido)prop-1-yn-1-yl)benzamido)propanoic acidwas obtained (45.00 mg, 55% yield) using the methyl2-(4-(3-(N-phenylpentaneamido)prop-1-yn-1-yl)benzamido)propanoate (86.50mg, 0.20 mmol) obtained in Example 43.

¹H NMR (CDCl₃, 500 MHz) δ 7.71 (2H, d, J=8.0 Hz, aromatic), 7.40 (7H, m,aromatic), 4.76 (1H, t, J=7.5 Hz and 14.5 Hz, CH), 4.72 (2H, s, CH₂),2.10 (2H, t, J=7.5 Hz and 15.5 Hz, CH₂), 1.57 (5H, m, CH₃, CH₂), 1.22(2H, m, CH₂), 0.80 (3H, t, J=7.5 Hz and 15.0 Hz, CH₃).

Example 45. Preparation of2-(4-(3-(N-(3-fluorophenyl)pentaneamido)prop-1-ynyl)phenoxy)acetic acid(LMT-1009) Step 1: Preparation of N-(3-fluorophenyl)pentaneamide

N-(3-fluorophenyl)pentaneamide was obtained (349.00 mg, 99% yield) bythe same method as described in Step 1 of Example 37 using3-fluoroaniline (200.00 mg, 1.79 mmol).

¹H NMR (CDCl₃, 500 MHz) δ 8.10 (1H, s), 7.51 (1H, d, J=11.0 Hz), 7.20(2H, m), 6.79 (1H, m), 2.36 (2H, t), 1.68 (2H, m), 1.36 (2H, m), 0.91(3H, t)

Step 2: Preparation of N-(3-fluorophenyl)-N-(prop-2-ynyl)pentaneamide

The N-(3-fluorophenyl)pentaneamide (400.00 mg, 2.05 mmol) obtained inStep 1, potassium hydroxide (230.61 mg, 4.11 mmol), and tetrabutylammonium iodide (37.87 mg, 0.20 mmol) were dissolved in tetrahydrofuran(20.00 ml), and stirred for 20 minutes. Propargyl bromide (0.19 ml, 2.30mmol) was added to the mixture at the same temperature, and stirred for20 hours. The reaction solution was concentrated under reduced pressure,and the obtained residue was diluted with ethyl acetate and washed withwater and brine. An organic solvent layer was collected, dehydrated withanhydrous magnesium sulfate (MgSO₄), filtered, and then concentratedunder reduced pressure. The concentrate was purified by silica gelcolumn chromatography (Hex:EA=10:1), thereby obtainingN-(3-fluorophenyl)-N-(prop-2-ynyl)pentaneamide (450.00 mg, 94% yield).

1H NMR (CDCl₃, 500 MHz) δ 7.42 (1H, m), 7.11 (2H, m), 7.20 (1H, d, J=9.0Hz), 4.46 (2H, s), 2.07 (2H, t), 1.56 (2H, m), 1.22 (2H, m), 0.82 (3H,t)

Step 3: Preparation ofN-(3-fluorophenyl)-N-(3-(4-hydroxyphenyl)prop-2-ynyl)pentaneamide

N-(3-fluorophenyl)-N-(3-(4-hydroxyphenyl)prop-2-ynyl)pentaneamide wasobtained (128.00 mg, 67.8% yield) using theN-(3-fluorophenyl)-N-(prop-2-ynyl)pentaneamide (270.00 mg, 1.16 mmol)obtained in Step 2 and 4-iodophenol (127.60 mg, 0.58 mmol).

¹H NMR (CDCl₃, 500 MHz) δ 7.51 (1H, m), 7.21 (5H, m), 6.71 (2H, d, J=9.0Hz), 7.65 (2H, s), 2.13 (2H, t), 1.54 (2H, m), 1.26 (2H, m), 0.82 (3H,t)

Step 4: Preparation of ethyl2-(4-(3-(N-(3-fluorophenyl)pentaneamido)prop-1-ynyl)phenoxy)acetate

Ethyl2-(4-(3-(N-(3-fluorophenyl)pentaneamido)prop-1-ynyl)phenoxy)acetate wasobtained (113.00 mg, 74% yield) using theN-(3-fluorophenyl)-N-(3-(4-hydroxyphenyl)prop-2-ynyl)pentaneamide(120.00 mg, 0.37 mmol) obtained in Step 3 and potassium carbonate(153.41 mg, 1.11 mmol).

¹H NMR (CDCl₃, 500 MHz) δ 7.42 (1H, m), 7.28 (2H, d, J=9.0 Hz), 7.09(3H, m), 6.81 (2H, d, J=9.0 Hz), 4.68 (2H, s), 4.61 (2H, s), 4.27 (2H,m), 2.05 (2H, t), 1.58 (2H, m), 1.26 (5H, m), 0.82 (3H, t)

Step 5: Preparation of2-(4-(3-(N-(3-fluorophenyl)pentaneamido)prop-1-ynyl)phenoxy)acetic acid

The ethyl2-(4-(3-(N-(3-fluorophenyl)pentaneamido)prop-1-ynyl)phenoxy)acetate(100.00 mg, 0.24 mmol) obtained in Step 4 was dissolved in ethanol (9.00ml), and stirred for 5 minutes. 2M sodium hydroxide (0.30 ml) was addedto the mixture, heated at 80° C., refluxed, and stirred for 3 hours. Thereaction solution was cooled at room temperature and concentrated underreduced pressure, and the obtained residue was diluted with ethylacetate and washed with water and brine. An organic solvent layer wascollected, dehydrated with anhydrous magnesium sulfate (MgSO₄),filtered, and then concentrated under reduced pressure. The concentratewas purified by ODS column chromatography (MeOD:H2O=2:1), therebyobtaining2-(4-(3-(N-(3-fluorophenyl)pentaneamido)prop-1-ynyl)phenoxy)acetic acid(15.00 mg, 16% yield)

¹H NMR (CDCl₃, 500 MHz) δ 7.52 (1H, d, J=7.0 Hz), 7.24 (5H, m), 6.88(2H, d, J=8.5 Hz), 4.68 (2H, s), 4.66 (2H, s), 2.14 (2H, t), 1.55 (2H,m), 1.24 (2H, m), 0.83 (3H, t)

Example 46. Preparation of2-(4-(3-(N-(4-fluorophenyl)pentaneamido)prop-1-ynyl)phenoxy)acetic acid(LMT-1010) Step 1: Preparation of N-(4-fluorophenyl)pentaneamide

N-(4-fluorophenyl)pentaneamide was obtained (870.00 mg, 99% yield) bythe same method as described in Step 1 of Example 37 using4-fluoroaniline (500.00 mg, 4.49 mmol) and valeroyl chloride (1.10 ml,8.99 mmol).

¹H NMR (CDCl₃, 400 MHz) δ 8.16 (1H, br), 7.48-7.45 (2H, m), 6.97-6.94(2H, m), 2.34-2.31 (2H, t), 1.68-1.65 (2H, m), 1.38-1.33 (2H, m),0.92-0.89 (3H, t).

Step 2: Preparation of N-(4-fluorophenyl)-N-(prop-2-ynyl)pentaneamide

N-(4-fluorophenyl)-N-(prop-2-ynyl)pentaneamide was obtained (347.60 mg,57% yield) by the same method as described in Step 2 of Example 45 usingthe N-(4-fluorophenyl)pentaneamide (500.00 mg, 2.56 mmol) obtained inStep 1 and propargyl bromide (0.24 ml, 2.81 mmol).

¹H NMR (CDCl₃, 500 MHz) δ 7.27 (2H, m), 7.14 (2H, m), 4.46 (2H, s), 2.04(2H, t), 1.55 (2H, m), 1.21 (2H, m), 0.81 (3H, t)

Step 3: Preparation ofN-(4-fluorophenyl)-N-(3-(4-hydroxyphenyl)prop-2-ynyl)pentaneamide

N-(4-fluorophenyl)-N-(3-(4-hydroxyphenyl)prop-2-ynyl)pentaneamide wasobtained (117.00 mg, 56% yield) using theN-(4-fluorophenyl)-N-(prop-2-ynyl)pentaneamide (300.00 mg, 1.29 mmol)obtained in Step 3 and 4-iodophenol (140.80 mg, 0.64 mmol).

¹H NMR (CDCl₃, 500 MHz) δ 7.22 (6H, m), 6.84 (2H, d, J=8.0 Hz), 4.65(2H, s), 2.06 (2H, t), 1.55 (2H, m), 1.21 (2H, m), 0.79 (3H, t)

Step 4: Preparation of ethyl2-(4-(3-(N-(4-fluorophenyl)pentaneamido)prop-1-ynyl)phenoxy)acetate

Ethyl2-(4-(3-(N-(4-fluorophenyl)pentaneamido)prop-1-ynyl)phenoxy)acetate wasobtained (113.00 mg, 81% yield) by the same method as described in Step1 of Example 27 using theN-(4-fluorophenyl)-N-(3-(4-hydroxyphenyl)prop-2-ynyl)pentaneamide(110.00 mg, 0.34 mmol) obtained in Step 3 and ethyl bromoacetate (0.04ml, 0.37 mmol).

¹H NMR (CDCl₃, 500 MHz) δ 7.21 (6H, m), 6.79 (2H, d, J=9.0 Hz), 4.65(2H, s), 4.59 (2H, s), 4.24 (2H, m), 2.01 (2H, t), 1.52 (2H, m), 1.21(5H, m), 0.79 (3H, t)

Step 5: Preparation of2-(4-(3-(N-(4-fluorophenyl)pentaneamido)prop-1-ynyl)phenoxy)acetic acid

2-(4-(3-(N-(4-fluorophenyl)pentaneamido)prop-1-ynyl)phenoxy)acetic acidwas obtained (25.00 mg, 27% yield) by the same method as described inStep 5 of Example 28 using the ethyl2-(4-(3-(N-(4-fluorophenyl)pentaneamido)prop-1-ynyl)phenoxy)acetate(100.00 mg, 0.24 mmol) obtained in Step 4.

¹H NMR (CDCl₃, 500 MHz) δ 7.41 (2H, m), 7.26 (4H, m), 6.88 (2H, d, J=7.0Hz), 4.66 (2H, s), 4.60 (2H, s), 2.10 (2H, t), 1.52 (2H, m), 1.23 (2H,m), 0.82 (3H, t)

EXPERIMENTAL EXAMPLES Experimental Example 1. Preparation ofBLT2-Expressing Cells or BLT2-Nonexpressing Cells

For this experiment, BLT2-nonexpressing cells and BLT2-expressing cells(CHO-BLT2 cells) were prepared by the following method.

CHO cells were obtained from Korean Cell Line Bank (KCLB, 10061), andcultured in an RPMI 1640 medium (Invitrogen) containing 10% fetal bovineserum (FBS; Life Technologies, Inc.), penicillin (50 units/mL) and anantibiotic antimycotic solution (Life Technologies, Inc.) at 37° C.under a 5% CO₂ condition. The cells were split for 3 days usingTrypsin-EDTA, maintained in a growth phase, washed withphosphate-buffered saline (PBS; 137 mM NaCl, 2.7 mM KCl, 10 mM Na₂HPO₄,2 mM KH₂PO₄), and then added to a new medium, thereby preparingBLT2-nonexpressing cells.

In addition, to prepare stable CHO/BLT2 clones, CHO-K1 cells weretransformed with pcDNA3-long form BLT2 encoding HA-tagged human BLT2,and selected with 0.4 mg/ml of G418 (Invitrogen, Carlsbad, Calif., USA).To screen BLT2 expression, the selected clones were analyzed by RT-PCRusing a human-specific BLT2 primer, and representative clones used forthe experiment were BLT2-expressing cells (CHO-BLT2 cells).

Experimental Example 2. Confirmation of Inhibitory Effect on Growth ofBLT2-Expressing Cells

Cell viability according to treatment of the compounds prepared in theexamples were measured by a3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT)method.

More specifically, 1×10⁴ each of the BLT2-nonexpressing cells(CHO-pcDNA3.1 cells) and BLT2-expressing cells (CHO-BLT2 cells), whichwere prepared in Experimental Example 1, were dispensed in a 96-mmculture dish, and cultured for 24 hours. Afterward, the culture mediumwas removed, a serum-free RPMI medium was added, and after two hours,the cells were pre-treated with each of the compound prepared in one ofthe examples (10 μM), 10 μM DMSO (compound solvent) as a control, and 10μM1-[5-ethyl-2-hydroxy-4-[[6-methyl-6-(1H-tetrazol-5-yl)heptyl]oxy]phenyl]-ethanone(LY255283; Cayman) as a positive control for 1 hour. Subsequently, aftertreatment of LTB₄ (300 nM), the cells were cultured for 24 hours. 20 μLof an MTT solution (5 mg/mL, Sigma-Aldrich) was added to each well, thecells were cultured in a humid CO₂ incubator at 37° C. for 4 hours, asupernatant was removed, and 200 μL of DMSO was added to each well todissolve insoluble violet formazan crystals. Absorbance was measuredusing a microplate reader (Molecular Devices, Sunnyvale, Calif.) at 550nm, and the measurement was repeated three times.

As a result, as shown in FIGS. 1A to 1E, when BLT2-expressing cells(CHO-BLT2 cells) were treated with LTB₄ (300 nM), which is a ligand ofBLT2 (DMSO+) were compared with BLT2-expressing cells (CHO-BLT2 cells)treated with ethanol (DMSO−), cell growth increased 20% to 35%, and whenBLT2-expressing cells (CHO-BLT2 cells) pre-treated with the positivecontrol LY255283, were compared with those treated with the controlDMSO, approximately 90% cell growth was exhibited, and therefore, it wasconfirmed that the inhibitory effect on cell growth was exhibited by thetreatment of the compounds of the examples. Specifically, when acompound of the present invention (LMT-692, LMT-694, LMT-696 orLMT-1013) was pre-treated at 10 μM, compared with the control DMSO,88.0%, 16.7%, 56.6% or 96.3% cell growth was exhibited, respectively,and thus the growth inhibitory effect was confirmed. Likewise, LMT-837(65%), LMT-841 (60%), LMT-842 (70%), LMT-883 (99%), LMT-886 (99%),LMT-1016 (99%), LMT-1018 (71.6%), and LMT-1019 (99%) compounds alsoshowed the growth inhibitory effect.

The experimental results show that the compounds of the presentinvention (LMT-692, LMT-696, LMT-837, LMT-841, LMT-842, LMT-883,LMT-886, LMT-1013, LMT-1016, LMT-1018, and LMT-1019) can inhibitBLT2-induced cell growth with very excellent efficiency, and thecompounds may be used as pharmaceutical components (BLT2-blockingpharmacological molecules) that can be used as therapeutic agents forinhibiting cancer, asthma or different types of BLT2-associatedinflammatory diseases.

Experimental Example 3. Confirmation of LTB₄-Induced BLT2-DependentChemotactic Motility Inhibitory Effect

Chemotactic motility was analyzed using a Transwell chamber including apolycarbonate filter (8-μm pore size, Corning Costar) with a 6.5-mmdiameter. Specifically, the lower surface of the filter was coated with10 μg/mL fibronectin in a serum-free RPMI 1640 medium at 37° C. for 1hour. The experiment was performed by placing the filter dried andcoated with RPMI 1640 media containing various amounts of LTB₄ in thelower wells of the Transwell chamber, and loading CHO cells stablyexpressing BLT1 and BLT2 into the upper wells containing serum-free RPMI1640 media finally at 2×10⁴ cells/100 μL. To evaluate the effect ofinhibitors, the cells were pre-treated with each inhibitor for 30minutes before dispensing. After the cells were cultured at 37° C. in 5%CO₂ for 3 hours, the filters were fixed with methanol for 3 minutes, andstained with hematoxylin and eosin for 10 minutes. In the experiment,the cells were BLT2-expressing cells (CHO-BLT2 cells) andBLT1-expressing cells (CHO-BLT1 cells), and LY255283 and U75302 wereused as positive controls for each type of the cells, and BLT2 ligandLTB₄, (300 nM), BLT1 ligand LTB₄ (10 nM), and lysophosphatidic acid(LPA; 100 nM) were used as comparative controls. The chemotacticmotility was quantitatively analyzed by counting the cells on the lowerside of the filter under an optical microscope (magnification, 200×).For each analysis, 6 fields were subjected to counting, each sample wasanalyzed twice, and the analysis was repeated three times.

As a result, as shown in FIGS. 2A and 2B and Table 1 below, in theBLT2-expressing cells (CHO-BLT2 cells), as the concentrations of thecompound of the present invention (LMT-692 or LMT-696) was increased(10⁻⁴, 10⁻³, 10⁻², 10⁻¹, 1, 10 and 10²), the chemotactic motility of theCHO-BLT2 cells was inhibited under a serum-free condition, and the 50%inhibitory concentrations (IC₅₀) of the LMT-692 and LMT-696 compoundswere 7.566 μM and 2.003 μM, respectively.

TABLE 1 IC50, μM receptor LTB₄, nM LMT-692 LMT-696 BLT2 300 7.566 2.003

In addition, as shown in Table 2 below, it was confirmed that, in theBLT2-expressing cells (CHO-BLT2 cells), as the concentration of thecompound of the present invention LMT-1013 is increased, the chemotacticmotility of the CHO-BLT2 cells was inhibited under a serum-freecondition, and the IC₅₀ of the LMT-1013 compound was 62.35 nM.

Likewise, it was confirmed that, in the BLT1-expressing cells (CHO-BLT1cells), as the concentration of the compound of the present inventionLMT-1013 was increased, the chemotactic motility of the CHO-BLT2 cellswas inhibited under a serum-free condition, and the IC₅₀ of the LMT-1013compound was 10 μM or more.

TABLE 2 IC50, nM Receptor LTB₄, nM LMT-1013 BLT1 10 >10 μM BLT2 30062.38

In addition, as shown in FIGS. 3A and 3B, when the BLT2-expressing cells(CHO-BLT2 cells) were treated with the BLT2 ligand LTB₄ (300 nM)(DMSO+), compared with those treated with ethanol (DMSO−), cellchemotactic motility increased 2.4-fold, and the cells pre-treated withLY255283 used as a positive control (10 μM) exhibited 90% chemotacticmotility as compared with the cells treated with the ligand LTB₄.Likewise, it was confirmed that, when the BLT1-expressing cells(CHO-BLT1 cells) were treated with the ligand LTB₄ (10 nM) (DMSO+),compared with those treated with ethanol (DMSO−), the cell chemotacticmotility was increased 2.2-fold, and the cells pre-treated with U75302used as a positive control (10 μM) exhibited 90% chemotactic motility ascompared with those treated with the ligand LTB₄. However, it wasconfirmed that, when the BLT2-expressing cells were pre-treated with thecompound of the present invention (LMT-692, LMT-694 or LMT-696) at 10μM, compared with those treated with the ligand LTB₄ (DMSO+), thechemotactic motility was 66%, 90% or 70.3% inhibited, respectively, butthe BLT1-expressing cells (CHO-BLT1 cells), compared with the ligandLTB₄ (DMSO+), did not exhibit the inhibitory effect on chemotacticmotility.

The results show that, in the cells in which BLT2 was stably expressed(CHO-BLT2 cells), chemotactic motility was increased due to LTB₄stimulus, the compound of the present invention (LMT-692, LMT-696, orLMT-1013) may considerably inhibit chemotactic motility, and thus can beused as a pharmaceutical component to inhibit LTB₄-inducedBLT2-dependent chemotactic motility.

Experimental Example 4. Confirmation of LTB₄ and BLT2 Binding InhibitoryEffect

The inhibition of LTB₄ and BLT2 binding (ligand binding affinity) wasanalyzed using radioactive tritium (³H)-labeled LTB₄ ([³H]LTB₄, ARC;specific activity 160.0 Ci/mmol). After 2×10⁶ of CHO-BLT2 cells wereplated into a 100-mm culture dish and cultured for 48 hours, anexperimental method was carried out as follows: Collected cells weretreated using a homogenizer a total of five times for 1 minute each toseparate proteins of the cell membrane. Afterward, the cells weresubjected to centrifugation at 4° C. and 45,000 rpm for 40 minutes toonly collect the proteins of the cell membrane, and thereby, a proteinconcentration of 40 μg/45 μL was quantified. When a BLT2-containing cellmembrane proteins which were quantified in the same manner was treatedwith the same amount of [³H]LTB₄ (5 nM), and then a differentconcentration (10⁻⁹, 10⁻⁸, 10⁻⁷, 10⁻⁶ or 10⁻⁵ M) of a compound, a degreeof inhibiting the tritium-labeled LTB₄ and BLT2 binding was measuredusing a Hidex 300sL liquid scintillation counter.

As a result, as shown in FIGS. 4A and 4B, it was confirmed that, in theBLT2-expressing cells (CHO-BLT2 cells), as the concentration of thecompound of the present invention (LMT-696 or LMT-1013) was increased(10⁻⁹, 10⁻⁸, 10⁻⁷, 10⁻⁶ and 10⁻⁵M), LTB₄ and BLT2 binding was inhibited,and the IC₅₀ of the LMT-696 and LMT-1013 compounds was 5.6 nM and 30.74nM, respectively.

Experimental Example 5. Confirmation of Anticancer Effect Due to BLT2Inhibition

The inventors have reported from previous research that BLT2 regulatesthe generation of intracellular reactive oxygen species (ROS) and acytokine interleukin-8 (IL-8) in breast cancer cells such as MDA-MB-231and MDA-MB-453 cells, resulting in the control of the invasion andmetastasis of cancer cells. Accordingly, it was confirmed that thegeneration of ROS and IL-8 expression were inhibited according to thetreatment of the compound of the present invention in MDA-MB-231 andMDA-MB-453 breast cancer cells.

5-1. Preparation of Breast Cancer Cells

The breast cancer cells such as the MDA-MB-231 cells were obtained fromKorean Cell Line Bank (Seoul, Korea), and the MDA-MB-435 cells wereprovided by J. H. Lee (Asan Medical Center, Seoul, Korea). These cellswere cultured in an RPMI 1640 medium (Invitrogen) containing 10% FBS(Life Technologies, Inc.), 1% penicillin (50 units/mL), and anantibiotic antimycotic solution (Life Technologies, Inc.) at 37° C.under a 5% CO₂ condition.

5-2. Confirmation of Inhibitory Effect on Intracellular ROS Generation

Intracellular ROS (H₂O₂) generated according to the treatment of thecompound of the present invention (LMT-696) was measured as a functionof DCF fluorescence. Specifically, before ROS measurement, 2×10⁵ cellswere grown in 60-mm wells, and cultured in a FBS-supplemented RPMI 1640medium for 24 hours. To evaluate the effect of the compound of thepresent invention, the cells were treated with the compound (LMT-696)for 30 minutes. To measure the intracellular ROS, the cells werecultured with 20 μM of a H₂O₂-sensitive fluorescent material such asH₂DCFDA [Molecular Probes (Eugene, Oreg.)] at 37° C. in a dark andhumidified CO₂ incubator for 20 minutes. The H₂DCFDA was hydrolyzed toDCF in the cells, and oxidized to DCF exhibiting high fluorescence inthe presence of H₂O₂, and thus the ROS amount was measured using as suchproperty. In addition, to confirm the ROS generation using a detector,the cells were harvested using trypsin-EDTA, and resuspended inserum-free RPMI 1640 without phenol red. A DCF fluorescent degree wasmeasured with excitation and emission wavelengths at 488 and 530 nm,respectively, using a FACS Calibur flow cytometer (Becton Dickinson,N.J.).

As a result, as shown in FIGS. 5A and 5B, it was confirmed that, whentreated with the compound of the present invention (LMT-696), the breastcancer cells such as the MDA-MB-231 and MDA-MB-435 cells exhibitedsignificant inhibition of the ROS generation.

5-3. Confirmation of Inhibitory Effect on IL-8 Expression

To confirm the IL-8 expression according to the treatment of thecompound of the present invention, total RNA was isolated from cellsusing Easy Blue (Intron, Sungnam, Korea), and quantified by absorbanceat 260 nm. Complementary DNA (cDNA) was synthesized with the RNA (1.25μg) through reverse transcription using a polymerase chain reaction(PCR) technique. An expression level was determined using primersspecifically binding to IL-8 and glyceraldehyde-3-phosphatedehydrogenase (GAPDH).

As a result, as shown in FIGS. 6A and 6B, it was confirmed that, whentreated with the compound of the present invention (LMT-696), the breastcancer cells such as the MDA-MB-231 and MDA-MB-435 cells exhibited asignificantly inhibited IL-8 expression level.

5-4. Confirmation of Inhibitory Effect on Invasion of Breast CancerCells

To detect the invasion of breast cancer cells according to the treatmentof the compound of the present invention, BioCoat Matrigel InvasionChambers (BD Biosciences, Bedford, Mass.) were used. 5×10⁴ of the breastcancer cells were harvested with trypsin-EDTA, resuspended in 0.5%serum-containing RPMI 1640, and transferred to Matrigel inserts. Fivepercent serum-containing RPMI 1640 was added to the lower chamber, andthe cells were cultured at 37° C. for 36 hours. Each filter was fixedwith methanol for 3 minutes, and stained with hematoxylin and eosin for10 minutes. The invasiveness of the cancer cells was quantified by cellcounts on the lower side of the filter under an optical microscope(magnification, 200×). In each analysis, 6 fields were quantified. Eachsample was analyzed twice, and the analysis was repeated three times.

As a result, as shown in FIGS. 7A and 7B, when treated with the compoundof the present invention (LMT-696), it was confirmed that the cancercell invasion was inhibited 70% for the MDA-MB-231 cells, and inhibited56% for the MDA-MB-435 cells.

5-5. Confirmation of Inhibitory Effect on Metastasis of Breast CancerCells

An experiment for the metastasis of breast cancer cells according to thetreatment of the compound of the present invention was approved by theEthics Committee of Korea University, and all experimental animals usedin this experiment were treated according to the approved guidelines ofthe Korea University Animal Care and Use Committee. Six-week-old femalenude mice (Charles River, Wilmington, Mass.) were injected with cancercells to confirm cancer cell metastasis. The breast cancer cells werepre-treated with the compound of the present invention (LMT-696, 10 μM),LY255283, U75302 and DMSO, and 24 hours later, harvested withtrypsin-EDTA, resuspended in PBS, and then 2×10⁶ of the breast cancercells were intraperitoneally injected into mice anesthetized withzoletil (50 mg/kg). After five days, the compound of the presentinvention (LMT-696; 2.5 mg/kg), LY255283 (2.5 mg/kg), U75302 (0.25mg/kg) and DMSO were injected intraperitoneally three times every fivedays. At 15 weeks after the injection of the breast cancer cells, themice were dissected to observe the cancer cell metastasis.

As a result, as shown in FIGS. 8, 9A and 9B, it was confirmed that themetastasis of the cancer cells (MDA-MB-231) was inhibited 40% by thetreatment of the compound of the present invention (LMT-696), wasinhibited 36% by the treatment of a positive control LY255283, ascompared with the control, and was not inhibited by the treatment ofU75302.

The results show that the compound of the present invention (LMT-696)can inhibit the generation of intracellular ROS and IL-8 of the cancercells, and thus inhibit the invasiveness and metastasis of the cancercells, and therefore the compound can be used as a pharmaceuticalcomponent having excellent anticancer efficiency.

Experimental Example 6. Confirmation of Anti-Asthma Effect by LBT2Inhibition

Mast cells play a pivotal role in the initial reaction to asthma, andwhen an allergen enters the body from the outside through an airway, themast cells are activated, thereby secreting various cytokines(interleukin-4 and interleukin-13). Due to the cytokines, the influx ofinflammatory cells, the generation of mucus and the airway contractionoccur. The inventors used 7-week-old (18 to 20 g) female BALB/c miceprovided by Orient (Seoungnam, Korea) for the experiment to confirm theantiasthma effect, and then the asthma was induced in the mice. On thefirst and 14^(th) days, 2.5 mg of an adjuvant, aluminum hydroxide gel(alum; Pierce, Rockford, Ill.) was included in 20 mg of ovalbumin (OVA)to intraperitoneally sensitize female C57BL/6 mice. On the 21^(st),22^(nd) and 23^(rd) days of two initial sensitizations, 1% OVA wassprayed into the mice using an ultrasonic nebulizer. The compound of thepresent invention (LMT-696; 5 mg/kg), LY255283 (5 mg/kg, Cayman) or DMSOwas intraperitoneally injected at the 1 hour before the 1% OVA spraying.On the 24^(th) day of the initial sensitization, airwayhyperresponsiveness (AHR) was detected, and on the 25^(th) day, the micewere dissected to observe asthma phenotypes, for example, inflammatorycytokine IL-4 expression and the influx of inflammatory cells(neutrophils). In the case of lipopolysaccharide (LPS)-induced severeasthma animal models, on the 0, 1^(st), 2^(nd) and 7^(th) days, 75 μg ofOVA and 1 mg of LPS were intranasally injected into Balb/c mice forsensitization. On the 14^(th), 15^(th), 21^(th) and 22^(th) days, 50 μgof OVA was injected into the nose for a challenge. The compound of thepresent invention (LMT-1013) (1, 3, 10 or 30 mg/kg), montelukast (10mg/kg, DRS) or a control buffer (10% DMA, 5% Tween 80, 85% brine) weretreated one hour before the challenge by injecting 50 μg of OVA into thenose. On the 23^(rd) day of the initial sensitization, AHR was detected,and on the 24^(th) day, the mice were dissected to observe a severeasthma phenotype, for example, the influx of the inflammatory cells(neutrophils). In addition, the AHR detection was performed after anairway constrictor, methacholine (6.25 to 50 mg/ml depending onconditions), was administered to the mice. The administration of theairway constrictor was performed by spraying through an inlet of thechamber using an ultrasonic nebulizer for 3 minutes. The AHR wasanalyzed using an enhanced pause as the indicator of the asthmaphenomenon. Bronchoalveolar lavage fluid cell counts were quantified bycounting cells under an optical microscope (magnification, 200×). Ineach analysis, 4 fields were subjected to counting, each sample wasanalyzed twice, and the analysis was repeated three times.

In addition, as shown in FIGS. 10 and 11, it was confirmed that, whenpre-treated with a positive control, LY255283, at 10 μM, the AHR of micein which severe asthma was induced by administering 50 mg/ml of anairway constrictor was reduced by 69.2%, and the generation of IL-4 inthe cells isolated from the abdominal cavity of the mice was reduced by67.2%. Further, when pre-treated with the compound of the presentinvention (LMT-696) at 10 μM, the AHR of mice in which severe asthma wasinduced by administering 50 mg/ml of an airway constrictor was reducedby 70%, and the generation of IL-4 of cells isolated from the abdominalcavity of the mice was reduced by 70%.

In addition, as shown in FIG. 12, in the asthma-induced mice (OVA+LPS),compared with mice in which asthma was not induced (Normal), AHR wasincreased 13-fold, and it was confirmed that, when pre-treated with thecompound of the present invention (LMT-1013) at 1, 3, 10 and 30 mg/kg,the asthma-induced mice (OVA+LPS), compared with mice to which 50 mg/mlof the airway constrictor was administered, AHR was reduced 48.6%,52.9%, 83.2%, and 87.3%, respectively. On the contrary, it was confirmedthat, compared with the mice to which 50 mg/ml of the airway constrictorwas administered, when pre-treated with a comparative material,montelukast, at 10 mg/kg, the AHR of the asthma-induced mice (OVA+LPS)was reduced 64%.

Further, as shown in FIGS. 13A, 13B and 13C, it was confirmed that, whenthe compound of the present invention (LMT-1013) was pre-treated at 1and 10 mg/kg, in the asthma-induced mice (OVA+LPS), total cells andneutrophils entering the abdominal cavity of the mice were reduced, andparticularly, the immune cells, that is, the neutrophils, were reduced51.6% and 90.3%, respectively.

Furthermore, as shown in FIGS. 14A and 14B, it was confirmed that, whenthe compound of the present invention (LMT-1013) was pre-treated at 1,3, 10, and 30 mg/kg, in the asthma-induced mice (OVA+LPS), total cellsand neutrophils entering the abdominal cavity of the mice were reduced,and particularly, the neutrophils were reduced by 42.2%, 48.8%, 71.8%,and 88.3%, respectively. On the contrary, it was confirmed that, whenthe comparative material montelukast was pre-treated at 10 mg/kg, theimmune cells, that is, the neutrophils, entering the abdominal cavity ofthe mice were not reduced.

The results showed that the compounds of the present invention (LMT-696and LMT-1013) inhibited AHR in asthma animal models, the compoundLMT-696 inhibited the generation of an inflammatory cytokine IL-4, andthe compound LMT-1013 inhibited the influx of immune cells into theabdominal cavity, resulting in the alleviation of the symptoms ofasthma, and therefore these compounds can be used as a pharmaceuticalcomponent having an antiasthma effect.

It should be understood by those of ordinary skill in the art that theabove description of the present invention is exemplary, and theexemplary embodiments disclosed herein can be easily modified into otherspecific forms without departing from the technical spirit or essentialfeatures of the present invention. Therefore, the exemplary embodimentsdescribed above should be interpreted as illustrative and not limited inany aspect.

INDUSTRIAL AVAILABILITY

The present invention relates to a novel compound having BLT2 inhibitoryactivity and a pharmaceutical composition for preventing or treating aninflammatory disease, which includes the compound. The inventorsidentified a novel compound containing BTL2 inhibitory activity to solvethe problems of the conventional compounds that had been designed totreat an inflammatory disease; for example, the instability in livingorganism and the difficulty on the mass production.

In addition, it was experimentally confirmed that the present novelcompound had an excellent effect on the enhancement of the cancer celldeath, on the inhibition of the metastasis and chemotactic mobility, andon the anti-asthma activity. Therefore, the present novel compound canbe used as a very effective pharmaceutical component for treating theinflammatory-related diseases.

1. A method for preventing or treating an inflammatory diseasecomprising: administering a pharmaceutical composition comprising acompound represented by Formula 1 or a pharmaceutically acceptable saltthereof to a subject in need thereof:

wherein, R₁ is C₁ to C₁₀ alkyl; R₂ is

wherein, R_(a) is

 or hydroxy; R_(b) is

R_(c) is

R_(d) is hydrogen or

 and R_(e) is

 and R₃ is hydrogen or fluorine.
 2. The method of claim 1, wherein R₁ isn-Butyl; R₂ is

wherein, R_(a) is

 or hydroxy; and R_(b) is


3. The method of claim 1, wherein the compound represented by Formula 1is selected from the group consisting of: tert-butyl4-(4-(3-(N-phenylpentaneamido)prop-1-ynyl)benzoyl)piperazine-1-carboxylate;N-phenyl-N-(3-(4-(piperazine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide;N-(3-(4-(4-methylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentaneamide;N-(3-(4-(4-ethylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentaneamide;N-(3-(4-(4-isopropylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentaneamide;N-(3-(4-(4-(2-hydroxyethyl)piperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentaneamide;N-(3-(4-(4-(cyclopropylmethyl)piperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentaneamide;N-(3-(4-(4-cyclohexylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentaneamide;N-(3-(4-(4-(cyclohexylmethyl)piperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentaneamide;N-(3-(4-(4-isobutylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentaneamide;N-phenyl-N-(3-(4-(4-(prop-2-ynyl)piperazine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide;N-(3-(4-(4-cyanopiperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentaneamide;tert-butyl4-(4-(3-(N-(3-fluorophenyl)pentaneamido)prop-1-ynyl)benzoyl)piperazine-1-carboxylate;N-(3-fluorophenyl)-N-(3-(4-(piperazine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide;N-(3-fluorophenyl)-N-(3-(4-(4-isopropylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide;tert-butyl4-(4-(3-(N-(4-fluorophenyl)pentaneamido)prop-1-ynyl)benzoyl)piperazine-1-carboxylate;N-(4-fluorophenyl)-N-(3-(4-(piperazine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide;N-(4-fluorophenyl)-N-(3-(4-(4-isopropylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide;N-(3-(4-(morpholine-4-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentaneamide;N-phenyl-N-(3-(4-(piperidine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide;N,N-diethyl-4-(3-(N-phenylpentaneamido)prop-1-ynyl)benzamide;N-phenyl-N-(3-(3-(piperazine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide;N-(3-(3-(4-methylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentaneamide;N-(3-(3-(4-isopropylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentaneamide;tert-butyl-4-(3-(3-(N-(4-fluorophenyl)pentaneamido)prop-1-ynyl)benzoyl)piperazine-1-carboxylate;N-(4-fluorophenyl)-N-(3-(3-(piperazine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide;N-(4-fluorophenyl)-N-(3-(3-(4-isopropylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide;N-(3-(4-hydroxyphenyl)prop-2-ynyl)-N-phenylpentaneamide;2-(4-(3-(N-phenylpentaneamido)prop-1-ynyl)phenoxy)acetic acid;tert-butyl4-(5-(3-((N-phenylpentaneamido)prop-1-yn-1-yl)picolinoyl)piperazine-1-carboxylate;N-phenyl-N-(3-(6-(piperazine-1-carbonyl)pyridine-3-yl)prop-2-yn-1-yl)pentaneamide;N-(3-(6-isopropylpiperazine-1-carbonyl)pyridine-3-yl)prop-2-yn-1-yl)pentaneamide;N,N-diethyl-4-(3-(N-(3-fluorophenyl)pentaneamido)prop-1-yn-1-yl)benzamide;N,N-diethyl-4-(3-(N-(4-fluorophenyl)pentaneamido)prop-1-yn-1-yl)benzamide;N-(3-(4-(N,N-diethylsulfamoyl)phenyl)prop-2-ynyl)-N-phenylpentaneamide;N-(3-(4-(N-isopropylsulfamoyl)phenyl)prop-2-ynyl)-N-phenylpentaneamide;tert-butyl 4-(3-(N-phenylpentaneamido)prop-1-yn-1-yl)benzoate;4-(3-(N-phenylpentaneamido)pro-1-yn-1-yl)benzoic acid;N-ethyl-4-(3-(N-phenylpentaneamido)prop-1-yn-1-yl)benzamide;N-(2-(diethylamino)ethyl)-4-(3-(N-phenylpentaneamido)prop-1-yn-1-yl)benzamide;ethyl 2-(4-(3-(N-phenylpentaneamido)prop-1-yn-1-yl)benzamido)acetate;2-(4-(3-(N-phenylpentaneamido)prop-1-yn-1-yl)benzamido)acetic acid;methyl2-(4-(3-(N-phenylpentaneamido)prop-1-yn-1-yl)benzamido)propanoate;2-(4-(3-(N-phenylpentaneamido)prop-1-yn-1-yl)benzamido)propionic acid;2-(4-(3-(N-(3-fluorophenyl)pentaneamido)prop-1-ynyl)phenoxy)acetic acid4-(3-(N-phenylpentaneamido)prop-1-yn-1-yl)benzoic acid;N-phenyl-N-(3-(4-(piperazine-1-carbonyl)phenyl)prop-2-ynyl)pentanamide;N-(2-(dimethylamino)ethyl)-4-(3-(N-phenylpentaneamido)prop-1-yn-1-yl)benzamide;2-(4-(3-(N-phenylpentaneamido)prop-1-yn-1-yl)benzamido)propanoic acid;and 2-(4-(3-(N-(4-fluorophenyl)pentaneamido)prop-1-ynyl)phenoxy)aceticacid.
 4. The method of claim 1, wherein the compound represented byFormula 1 is selected from the group consisting of:N-(3-(4-(4-methylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentaneamide;N-phenyl-N-(3-(4-(piperazine-1-carbonyl)phenyl)prop-2-ynyl)pentanamide;N-(3-(4-(4-isopropylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentaneamide;N-phenyl-N-(3-(3-(piperazine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide;N-(4-fluorophenyl)-N-(3-(4-(4-isopropylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide;N-(3-(3-(4-isopropylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentaneamide;N,N-diethyl-4-(3-(N-phenylpentaneamido)prop-1-ynyl)benzamide;N-(3-fluorophenyl)-N-(3-(4-(4-isopropylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)pentaneamide;4-(3-(N-phenylpentaneamido)prop-1-yn-1-yl)benzoic acid;N-(2-(dimethylamino)ethyl)-4-(3-(N-phenylpentaneamido)prop-1-yn-1-yl)benzamide;methyl2-(4-(3-(N-phenylpentaneamido)prop-1-yn-1-yl)benzamido)propanoate; and2-(4-(3-(N-phenylpentaneamido)prop-1-yn-1-yl)benzamido)propanoic acid.5. The method of claim 1, wherein the inflammatory disease is selectedfrom the group consisting of asthma, atherosclerosis, cancer, pruritus,rheumatoid arthritis and inflammatory enteropathy.
 6. The method ofclaim 1, wherein the pharmaceutical composition inhibits leukotriene B4receptor 2 (BLT2) activity.